KR20210021467A - Activatable interleukin-2 polypeptide and method of use thereof - Google Patents

Abstract

The present disclosure features a fusion protein that is a conditionally active variant of IL-2. In one aspect, the full-length polypeptide of the present invention has a reduced or minimal cytokine-receptor activation activity even when it contains a functional cytokine polypeptide. For example, upon activation of a blocking moiety, e.g., a steric blocking polypeptide, by cleavage of a linker that connects to a sequence for an active cytokine, the cytokine can bind to its receptor and affect signaling. have.

Description

Activatable interleukin-2 polypeptide and method of use thereof

Related application

This application is of U.S. Provisional Application 62/671,225 filed May 14, 2018, U.S. Provisional Application No. 62/756,504 filed November 6, 2018, and U.S. Provisional Application No. 62/756,507 filed November 6, 2018. Claim profit The entire teachings of this application are incorporated herein by reference.

Sequence list

This application contains a sequence listing, filed electronically in ASCII format and incorporated herein by reference in its entirety. The ASCII copy, created on May 14, 2019, is named 105365-0021_SL.txt and is 408,319 bytes in size.

The development of mature immunocompetent lymphocytes from less-entangled precursors, their subsequent antigen-induced immune responses, and inhibition of these and unwanted autoreactions are highly dependent and common γ-chain (γc) family (Rochman et al., 2009) and Il Regulated by cytokines (including interleukin-2 [IL-2], IL-4, IL-7, IL-9, IL-15 and IL-21) using receptors in family members including -12, 18 and 23 do. IL-2 is essential for thymic development of Treg cells and critically regulates several major aspects of mature peripheral Tregs and antigen-activated existing T cells. Due to its potent T cell growth factor activity in vitro, IL-2 has been studied extensively in part, as this activity offers a potential means to directly boost immunity, for example in cancer and AIDS-HIV patients. Because it provides targets to antagonize unwanted reactions, such as transplant rejection and autoimmune diseases. Although in vitro studies with IL-2 provided a strong basis for these studies, the function of IL-2 in vivo is clearly much more complex, as first described in IL-2-deficient mice, where rapid immunity is not lacking. Fatal autoimmune syndrome was observed (Sadlack et al., 1993, 1995). Similar observations were made later when the genes encoding IL-2Rα (Il2ra) and IL-2Rβ (Il2rb) were individually removed (Suzuki et al., 1995; Willerford et al., 1995).

The present invention is directed to conditionally active and/or targeted cytokines for use in the treatment of cancer and other diseases that rely on immune up or down regulation. For example, the antitumor activity of some cytokines is well known and described, and some cytokines have already been used therapeutically in humans. Cytokines such as interleukin-2 (IL-2) have shown benign antitumor activity in patients with various types of tumors, such as renal metastatic carcinoma, hair cell leukemia, Kaposi's sarcoma, melanoma, multiple myeloma, etc. Other cytokines such as IFNβ, tumor necrosis factor (TNF)α, TNFβ, IL-1, 4, 6, 12, 15 and CSF have shown specific antitumor activity against some types of tumors and are therefore subject to further study. .

summary

Therapeutic proteins, nucleic acids encoding proteins, and diseases such as proliferative diseases, neoplastic diseases, inflammatory diseases, immune diseases, autoimmune diseases, infectious diseases, viral diseases, allergic reactions, parasitic reactions, graft-versus-host diseases, etc. or Compositions and methods of using proteins and nucleic acids for the treatment of disorders are provided herein.

The present invention features a fusion protein that is a conditionally active variant of IL-2. In one aspect, the full-length polypeptide of the present invention has reduced or minimal IL-2-receptor activation activity even if it contains a functional cytokine polypeptide. For example, upon activation by cleavage of a linker connecting a blocking moiety, e.g., a steric blocking polypeptide, successively to an active cytokine, IL-2, or a functional fragment or mutein thereof, binds to its receptor It can affect signaling. If desired, the full-length polypeptide may also include blocking polypeptide moieties that provide additional advantageous properties. For example, the full-length polypeptide may also contain blocking polypeptide moieties that extend serum half-life and/or target the full-length polypeptide to the desired active site of IL-2. Alternatively, the full length fusion polypeptide may contain a serum half-life extending element and/or targeting domain that is distinct from the blocking polypeptide moiety. Preferably, the fusion protein contains at least one element or domain capable of extending the circulating half-life in vivo. Preferably, this element is enzymatically removed at the desired body location (e.g., protease cleavage in the tumor microenvironment), so that a payload molecule substantially similar to the naturally occurring payload molecule (e.g., IL2 or IFNa ) To restore the pharmacokinetic properties. Preferably, the fusion protein is targeted to the desired cell or tissue. Targeting as described herein is achieved through the action of a blocking polypeptide moiety that also binds to the desired target or through the targeting domain. Domains that recognize the target antigen on a desired target (eg, a tumor-specific antigen) can be attached to the cytokine through a cleavable or non-cleavable linker. When attached by a non-cleavable linker, the targeting domain can further help retain cytokines in the tumor and can be considered a retention domain. The targeting domain need not necessarily be linked directly to the payload molecule, but can be linked directly to other elements of the fusion protein. This is especially the case when the targeting domain is attached via a cleavable linker.

In one aspect there is provided a fusion polypeptide comprising an IL-2 polypeptide, or a functional fragment or mutein thereof, and a blocking moiety, such as a steric blocking domain. The blocking moiety is fused to the IL-2 polypeptide directly or through a linker, and by cleavage of the fusion polypeptide at or near or at the fusion site or linker (e.g., protease mediated cleavage) It can be isolated from the polypeptide. For example, when a cytokine polypeptide is fused to a blocking moiety through a linker containing a protease cleavage site, the cytokine polypeptide is released from the blocking moiety upon protease mediated cleavage of the linker and can bind to its receptor. . Linkers are designed to cleave at the site of desired cytokine activity, for example in the tumor microenvironment, to avoid off-target cytokine activity and reduce the overall toxicity of cytokine therapy.

The blocking moiety can also function as a serum half-life extending element. In some embodiments, the fusion polypeptide further comprises a separate serum half-life extending element. In some embodiments, the fusion polypeptide further comprises a targeting domain. In various embodiments, the serum half-life extending element is any branched or multi-armed polyethylene glycol (PEG), full-length human serum albumin (HSA) or fragment that preserves binding to FcRn, an Fc fragment, or directly to FcRn. Or a water-soluble polypeptide such as a nanobody that binds to human serum albumin.

In addition to the serum half-life extending element, the pharmaceutical compositions described herein preferably comprise at least one or more targeting domains that bind to one or more target antigens or one or more regions on a single target antigen. It is contemplated herein that the polypeptide constructs of the invention will be cleaved in the blood of a subject, for example at a disease-specific microenvironment or protease cleavage site and the targeting domain(s) will bind to the target antigen on the target cell. At least one target antigen is associated with and/or associated with a disease, disorder or condition. Exemplary target antigens are proliferative diseases, neoplastic diseases, inflammatory diseases, immunological disorders, autoimmune diseases, infectious diseases, viral diseases, allergic reactions, parasitic reactions, graft-versus-host diseases or host-versus-graft diseases. Includes those related to.

In some embodiments, the target antigen is a cell surface molecule such as a protein, lipid or polysaccharide. In some embodiments, the target antigen is a tumor cell, a virus infected cell, a bacterial infected cell, an damaged red blood cell, an arterial plaque cell, or a fibrous tissue cell.

The target antigen is optionally expressed on the surface of disease cells or tissues, such as tumors or cancer cells. Target antigens for tumors are fibroblast activating protein alpha (FAPa), trophoblast glycoprotein (5T4), tumor-associated calcium signal transducer 2 (Trop2), fibronectin EDB (EDB-FN), fibronectin EIIIB domain, CGS-2, EpCAM, EGFR, HER-2, HER-3, c-Met, FOLR1 and CEA. The pharmaceutical compositions disclosed herein also include proteins comprising two antigen binding domains that bind to two different target antigens known to be expressed in diseased cells or tissues. Exemplary pairs of antigen binding domains include, but are not limited to, EGFR/CEA, EpCAM/CEA and HER-2/HER-3.

In some embodiments, the targeting polypeptide independently comprises a scFv, a VH domain, a VL domain, a non-Ig domain, or a ligand that specifically binds a target antigen. In some embodiments, the targeting polypeptide specifically binds to a cell surface molecule. In some embodiments, the targeting polypeptide specifically binds to a tumor antigen. In some embodiments, the targeting polypeptide specifically and independently binds a tumor antigen selected from at least one of EpCAM, EGFR, HER-2, HER-3, cMet, CEA, and FOLR1. In some embodiments, the targeting polypeptide specifically and independently binds to two different antigens, wherein at least one of the antigens is a tumor antigen selected from EpCAM, EGFR, HER-2, HER-3, cMet, CEA and FOLR1. In some embodiments, the targeting polypeptide acts as a retention domain and is attached to a cytokine through a non-cleavable linker.

As described herein, cytokine blocking moieties can bind to IL-2 and thereby block activation of the IL-2 cognate receptor.

The present disclosure also relates to nucleic acids encoding conditionally active proteins described herein, such as DNA, RNA, mRNA, as well as vectors and host cells containing such nucleic acids.

The present disclosure also relates to conditionally active proteins, nucleic acids encoding conditionally active proteins, and pharmaceutical compositions containing vectors and host cells containing such nucleic acids. Typically, the pharmaceutical composition contains one or more physiologically acceptable carriers and/or excipients.

The present disclosure also provides an effective amount of a conditionally active protein, a nucleic acid encoding a conditionally active protein, a vector or host cell containing such nucleic acid, and a pharmaceutical composition of any of the foregoing, in need of treatment. It relates to a method of treatment comprising administering to a subject. Typically, the subject is a proliferative disease, neoplastic disease, inflammatory disease, immunological disorder, autoimmune disease, infectious disease, viral disease, allergic reaction, parasitic reaction, graft-versus-host disease or host-versus-graft disease. There is a risk of having or developing.

The present disclosure also provides a conditionally active protein, a nucleic acid encoding a conditionally active protein, a vector or host cell containing such a nucleic acid, and any of the foregoing for treating a subject in need of treatment. It relates to the use of pharmaceutical compositions. Typically the subject has a proliferative disease, a neoplastic disease, an inflammatory disease, an immunological disorder, an autoimmune disease, an infectious disease, a viral disease, an allergic reaction, a parasitic reaction, a graft-versus-host disease or a host-versus-graft disease. There is a risk of having or developing.

The present disclosure also provides for diseases such as proliferative diseases, neoplastic diseases, inflammatory diseases, immunological disorders, autoimmune diseases, infectious diseases, viral diseases, allergic reactions, parasitic reactions, graft-versus-host diseases or host-versus -Relates to the use of conditionally active proteins, nucleic acids encoding conditionally active proteins, vectors or host cells containing such nucleic acids for the manufacture of medicaments for the treatment of graft diseases.

1A is a schematic diagram illustrating a protease-activated cytokine or chemokine comprising a blocking moiety. The blocking moiety can optionally function as a serum half-life extension domain. To the left of the arrow, the figure shows that the cytokine is linked to a blocking moiety through a protease-cleavable linker, thus blocking its ability to bind to its receptor. The figure to the right of the arrow shows that in an inflammatory or tumor environment, the protease is cleaved at the protease-cleaving site on the linker, releasing the blocking moiety and allowing the cytokine to bind its receptor.
1B is a schematic diagram illustrating a protease-activated cytokine or chemokine in which HSA (blocking moiety) is directly bound to the cytokine or chemokine of interest, as a protease cleavage site between the HSA and the cytokine or chemokine of interest. . To the left of the arrow, the figure shows that the cytokine is linked to a blocking moiety through a protease-cleavable linker, thus blocking its ability to bind to its receptor. The figure to the right of the arrow shows that in an inflammatory or tumor environment, the protease is cleaved at the protease-cleaving site on the linker, releasing the blocking moiety and allowing the cytokine to bind its receptor.
1C is a schematic diagram illustrating a protease-activated cytokine or chemokine in which more than one HSA (blocking moiety) is directly bound to a molecule of interest. If desired, one or more HSAs may be linked to cytokines or chemokines through linkers, such as linkers containing protease cleavage sites. To the left of the arrow, the figure shows that the cytokine is linked to a blocking moiety through a protease-cleavable linker, thus blocking its ability to bind to its receptor. The figure to the right of the arrow shows that in an inflammatory or tumor environment, the protease is cleaved at the protease-cleaving site on the linker, releasing the blocking moiety and allowing the cytokine to bind its receptor. Cytokines now have similar pK properties compared to native cytokines (eg, have a short half-life).
1D is a schematic diagram illustrating a protease-activated cytokine or chemokine comprising one or more cytokines of the same or different types each linked to a binding domain via a protease-cleavable linker. To the left of the arrow, the figure shows that the cytokine is linked to a blocking moiety through a protease-cleavable linker, thus blocking its ability to bind to its receptor. The figure to the right of the arrow shows that in an inflammatory or tumor environment, the protease is cleaved at the protease-cleaving site on the linker, releasing the blocking moiety and allowing the cytokine to bind its receptor.
Figure 2 is a schematic diagram illustrating a protease-activated cytokine or chemokine comprising a cytokine or chemokine polypeptide, a blocking moiety and a serum half-life extension domain linked by at least one protease-cleavable linker. To the left of the arrow, the figure shows that the cytokine is linked to a blocking moiety through a protease-cleavable linker, thus blocking its ability to bind to its receptor. It is also bound to a separate half-life extending element, extending the half-life of the serum. The figure to the right of the arrow shows that in an inflammatory or tumor environment, the protease is cleaved at the protease-cleaving site on the linker, releasing serum half-life extending elements and blocking moieties and allowing cytokines to bind to their receptors. Cytokines now have similar pK properties compared to native cytokines (eg, short half-life).
3 is a schematic diagram illustrating a protease-activated cytokine or chemokine comprising a cytokine or chemokine polypeptide, a blocking moiety and a targeting domain linked by at least one protease-cleavable linker. To the left of the arrow, the figure shows that the cytokine is linked to the blocking moiety and targeting domain via a protease-cleavable linker, thus blocking its ability to bind its receptor. The figure to the right of the arrow shows that in the inflammation or tumor microenvironment, the protease is cleaved at the protease-cleaving site on the linker to release the targeting domain and blocking moiety and allow the cytokine to bind its receptor.
4A is a schematic diagram illustrating a protease-activated cytokine or chemokine comprising a cytokine or chemokine polypeptide, a blocking moiety, a targeting domain and a serum half-life extension domain linked by at least one protease-cleavable linker, wherein The cytokine polypeptide and targeting domain are linked by a protease-cleavable linker. To the left of the arrow, the figure shows that a is linked to the targeting domain, blocking moiety and half-life extending element via a protease-cleavable linker(s), thus blocking the ability to bind to its receptor. To the right of the arrow, the figure shows that in an inflammatory or tumor environment, the protease is cleaved at the protease-cleaving site on the linker(s) to release the half-life extending elements, targeting domains and blocking moieties and allow the cytokine to bind its receptor. Shows that. Cytokines now have similar pK properties compared to native cytokines (eg, short half-life).
Figure 4B is a schematic diagram illustrating a protease-activated cytokine or chemokine comprising a cytokine or chemokine polypeptide, a blocking moiety, a targeting domain and a serum half-life extension domain linked by at least one protease-cleavable linker. To the left of the arrow, the figure shows that the cytokine is linked to the targeting domain, blocking moiety and half-life extending element via a protease-cleavable linker(s), thus blocking the ability to bind to its receptor. The figure to the right of the arrow shows that in an inflammatory or tumor environment, the protease is cleaved at the protease-cleaving site on the linker(s) to release the half-life extending elements and blocking moieties and allow the cytokine to bind its receptor. . The targeting moiety remains bound to maintain cytokines in the tumor microenvironment. Cytokines now have similar pK properties compared to native cytokines (eg, short half-life).
5 is a schematic diagram illustrating the structure of the variable domain of an immunoglobulin molecule. The variable domains of the light and heavy chain immunoglobulin chains contain three hypervariable loops or complementarity-determining regions (CDRs). The three CDRs of the V domain (CDR1, CDR2, CDR3) are clustered at one end of the beta barrel. CDRs are loops connecting the beta strands BC, C'-C" and FG of the immunoglobulin fold, while the lower loop connecting the beta strands AB, CC', C"-D and EF of the immunoglobulin fold and the immunoglobulin fold The top loop connecting the DE strands is a non-CDR loop.
Figure 6 is a schematic diagram illustrating a protease-activated cytokine or chemokine comprising a cytokine or chemokine polypeptide, a blocking moiety that is a serum albumin binding domain (e.g., dAb) and a protease cleavable linker. In the illustrated example, the non-CDR loop of the serum albumin binding domain (eg, sdAb) can form a binding site for the cytokine IL-2. In this example, the binding site for serum albumin can be formed by the CDRs of the serum albumin binding domain.
7A-7H are a series of graphs showing the activity of exemplary IL-2 fusion proteins in the IL-2 dependent cytotoxic T lymphocyte cell line CTLL-2. Each graph represents the results of the IL-2 proliferation assay quantified by CellTiter-Glo® (Promega) luminescence-based cell viability assay. Each proliferation assay was performed with or without HSA (Figs. 7b, 7d, 7e, 7g) or without (Figs. 7a, 7c, 7f, 7h). Each fusion protein contained an anti-HSA binding agent, and both the uncleaved version and the MMP9 protease cleaved version of the fusion protein were used in each assay.
8A-8F are a series of graphs showing the activity of exemplary IL-2 fusion proteins in the IL-2 dependent cytotoxic T lymphocyte cell line CTLL-2. Each graph represents the results of IL-2 proliferation assay quantified by CellTiter-Glo (Promega) luminescence-based cell viability assay. The uncleaved version and the MMP9 protease cleaved version of the fusion protein were used for each assay.
9A-9Z are a series of graphs showing the activity of exemplary IL-2 fusion proteins in the IL-2 dependent cytotoxic T lymphocyte cell line CTLL-2. Each graph represents the results of IL-2 proliferation assay quantified by CellTiter-Glo (Promega) luminescence-based cell viability assay. The uncleaved version and the MMP9 protease cleaved version of the fusion protein were used for each assay.
10 shows the results of a protein cleavage assay as described in Example 3. Fusion protein ACP16 was run on SDS-PAGE gels in both cleaved and uncleaved forms. As can be seen from the gel, the cleavage was complete.
11 is a series of graphs depicting the results from the HEK-blue IL-2 reporter assay performed on IL-2 fusion protein and recombinant human IL2 (Rec hIL-2). The assay was performed based on the quantification of secreted alkaline phosphatase (SEAP) activity using the reagent QUANTI-Blue (InvivoGen).
12A and 12B are two graphs showing the analysis of ACP16 (12a) and ACP124 (12b) in the HEKBlue IL-2 reporter assay in the presence of HSA. The circle represents the activity of the uncut polypeptide and the square represents the activity of the truncated polypeptide. 12C is a graph showing the results of CTLL-2 proliferation analysis. CTLL2 cells (ATCC) were plated in suspension at a concentration of 500,000 cells/well in culture medium with or without 40 mg/ml human serum albumin (HSA) and a series of activatable hIL2 at _{37° C. and 5% CO 2 for 72 hours.} Stimulated with dilution. The activity of uncleaved and cleaved activatable ACP16 was tested. The cleaved activatable hIL2 was generated by incubation with active MMP9. Cellular activity was assessed using CellTiter-Glo (Promega) luminescence-based cell viability assay. Circles represent intact fusion proteins and squares represent protease-cleaved fusion proteins. Figure 12d
13 is three graphs showing results of analysis of ACP16 and ACP124 in a tumor xenograft model. 13A shows 4.4 μg ACP16 (square), 17 μg ACP16 (triangle), 70 μg ACP16 (lower triangle), 232 μg ACP16 (dark circle) and 12 μg wild type IL-2 (dotted line, triangle) and 36 μg wild type IL-2 (dotted line) as comparators. , Shows tumor volume over time in mice treated with diamonds Vehicle alone is represented by large open circles Data show tumors over time in a dose-dependent manner in mice treated with ACP16 at higher concentrations. Figure 13b shows tumor volumes over time in mice treated with 17 μg ACP124 (square), 70 μg ACP124 (triangle), 230 μg ACP124 (lower triangle) and 700 μg ACP124. Vehicle alone is large Figure 13c shows 17 μg ACP16 (triangle), 70 μg ACP16 (circle), 232 μg ACP16 (dark circle) and 17 μg ACP124 (dotted line, triangle) 70 μg ACP124 (dotted line, diamond), 230 μg ACP124 (dotted line) as a comparator. Diamond), the tumor volume over time in mice treated with the vehicle alone is indicated by a dark downward triangle Data is in a dose-dependent manner in mice treated with ACP16 rather than ACP124, tumor volume over time Indicates a decrease in
14A-C are a series of spaghetti plots showing the activity of fusion proteins in the MC38 mouse xenograft model corresponding to the data shown in FIG. 13. Each line in the plot is a single mouse.
15 is a graph showing tumor volume over time in a mouse xenograft model showing tumor growth in control mice (open circles) and AP16-treated mice (squares).
16 is a series of survival plots showing the survival of mice over time after treatment with a cleavable fusion protein. Figure 16A shows data for mice treated with vehicle alone (gray line), 17 μg ACP16 (dark line) and 1 μg ACP124 (dotted line). Figure 16B shows data for mice treated with vehicle alone (gray line), 70 μg ACP16 (dark line) and 70 μg ACP124 (dotted line). Figure 16C shows data for mice treated with vehicle alone (gray line), 232 μg ACP16 (dark line) and 230 μg ACP124 (dotted line). 16D shows data for mice treated with vehicle alone (grey line), 232 μg ACP16 (dark line) and 700 μg ACP124 (dotted line).
17 is a series of spaghetti plots showing the activity of fusion proteins in the MC38 mouse xenograft model. All groups of mice were given a total of 4 doses except for the maximum 3 doses of APC132, where lethal toxicity was detected after 1 week/2 doses. Vehicle alone (top), 17, 55, 70 and 230 μg ACP16 (top full row), 9, 38, 36 and 119 μg ACP132 (middle full row) and 13, 42, 54 and 177 μg ACP21 (bottom full row) are shown have. Each line in the plot represents an individual animal.
18-21 illustrate the properties of TriTac polypeptides acting as exemplary protease cleavable fusion proteins.

Methods and compositions for engineering and using constructs comprising inducible cytokines are disclosed herein. Cytokines are potent immune agents that make them considered promising therapeutics for oncology. However, it has been demonstrated that cytokines have a very narrow therapeutic range. Cytokines have a short serum half-life and are considered very potent. As a result, therapeutic administration of cytokines has caused undesirable systemic effects and toxicity. They are exacerbated by the need to administer large amounts of cytokines to achieve the desired levels of cytokines at the intended site of cytokine action (eg, tumor). Unfortunately, due to the inability to effectively target and control the biology of cytokines and their activity, cytokines have not achieved the clinical benefits expected in the treatment of tumors.

Fusion proteins are disclosed herein that overcome the toxicity and short half-life problems that severely limit the clinical use of cytokines in oncology. Fusion proteins contain cytokine polypeptides with receptor agonist activity. However, in the context of fusion proteins, cytokine receptor agonist activity is weakened and the circulating half-life is prolonged. Fusion proteins contain protease cleavage sites that are cleaved by proteases associated with the desired cytokine active site (eg, tumor), and are typically abundant or selectively present at the desired active site. Thus, the fusion protein is preferentially (or selectively) and efficiently cleaved at the desired active site to substantially limit cytokine activity to the desired active site, such as the tumor microenvironment. Protease cleavage at the desired active site, such as in the tumor microenvironment, releases a form of cytokine from the fusion protein that is much more active (typically at least about 100X more active than the fusion protein) as a cytokine receptor agonist than the fusion protein. The form of cytokines released upon cleavage of a fusion protein typically has a short half-life, which is often substantially similar to that of naturally occurring cytokines, further limiting cytokine activity to the tumor microenvironment. Even if the half-life of the fusion protein is prolonged, toxicity is dramatically reduced or eliminated because circulating fusion proteins are weakened and active cytokines target the tumor microenvironment. The fusion proteins described herein allow for the first time the administration of effective therapeutic doses of cytokines, allowing the treatment of tumors with the activity of cytokines that are substantially limited to the tumor microenvironment, the unwanted systemic effects of cytokines and It dramatically reduces or eliminates toxicity.

Unless otherwise defined, all technical terms, notations, and other scientific terms used herein are intended to have the meanings commonly understood by one of ordinary skill in the art to which this invention belongs. In some instances, terms having a generally understood meaning are defined herein for clarity and/or ease of reference, and the inclusion of such definitions herein necessarily indicates a difference from that generally understood in the art. Should not be interpreted as. The techniques and procedures described or referenced herein are generally well understood and, for example, Sambrook et al., Molecular Cloning: A Laboratory Manual 4th ed. (2012) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, using conventional methodologies such as the widely used molecular cloning methodology described. Suitably, procedures involving the use of commercially available kits and reagents are generally performed according to manufacturer-defined protocols and conditions unless otherwise specified.

“Cytokine” is a well-known technical term specifically referring to any class of immunomodulatory proteins (eg, interleukins or interferons) that are secreted by cells of the immune system and are modulators of the immune system. Cytokine polypeptides that can be used in the fusion proteins disclosed herein include transforming growth factors such as TGF-α and TGF-β (eg, TGFbeta1, TGFbeta2, TGFbeta3); Interferons such as interferon-α, interferon-β, interferon-γ, interferon-kappa and interferon-omega; Interleukin such as IL-1, IL-1α, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11 , IL-12, IL-13, IL-14, IL-15, IL-16, IL-17, IL-18, IL-21 and IL-25; Tumor necrosis factors such as tumor necrosis factor alpha and lymph toxins; Fragments of these polypeptides that activate chemokines (e.g. CXC motif chemokine 10 (CXCL10), CCL19, CCL20, CCL21) and granulocyte macrophage-colony stimulating factor (GM-CS), as well as cognate receptors for cytokines (I.e., functional fragments of the foregoing), but are not limited to these. "Chemokines" is a technical term referring to a family of any small cytokines that have the ability to induce directed chemosis in nearby reactive cells.

Cytokines are well known to have short serum half-lives, frequently of only a few minutes or hours. Even forms of cytokines with modified amino acid sequences that are intended to prolong serum half-life but retain receptor agonist activity typically also have short serum half-life. As used herein, a "short-half-life cytokine" refers to a serum half-life of less than 10 minutes, less than 15 minutes, less than 30 minutes, less than 60 minutes, less than 90 minutes, less than 120 minutes, less than 240 minutes, or less than 480 minutes. Likewise, it refers to cytokines with a substantially short half-life that circulate in the serum of a subject. As used herein, short half-life cytokines are cytokines whose sequence has not been modified to achieve a longer half-life than the normal half-life in the subject's body, and modifications intended to prolong serum half-life but retain receptor agonist activity. It includes a polypeptide having an amino acid sequence. Typically, short half-life cytokine polypeptides, such as IL-2 polypeptides, are serum comparable to naturally occurring IL-2, e.g., no more than 5, 4, 3 or 2 times that of naturally occurring IL-2. It has a half-life. This latter case does not imply the inclusion of the addition of heterologous protein domains such as true half-life extending elements such as serum albumin.

“Sortase” is a transpeptidase that modifies a protein by recognizing and cleaving a carboxyl-terminal classification signal inserted into or attached to a target protein or peptide. Sortase A catalyzes the cleavage of the LPXTG motif (SEQ ID NO: 125) (where X is any standard amino acid) between the Thr and Gly residues on the target protein and is an enzyme by transient attachment of the Thr residue to the active site Cys residue on the enzyme. -To form a thioacyl intermediate. In order to complete the transpeptization and generate a peptide-monomer conjugate, a biomolecule with an N-terminal nucleophilic group, typically an oligoglycine motif, attacks the intermediate to replace Sortase A and binds the two molecules.

As used herein, the term “stereoblocker” may be covalently linked to a cytokine polypeptide, for example in the form of a chimeric polypeptide (fusion protein), either directly or indirectly through another moiety such as a linker, or Otherwise, it refers to a polypeptide or polypeptide moiety that does not covalently bind to a cytokine polypeptide. Steric blocking agents can non-covalently bind cytokine polypeptides, for example via electrostatic, hydrophobic, ionic or hydrogen bonding. Steric blockers typically inhibit or block the activity of cytokine moieties due to their proximity and comparative size to the cytokine moiety. Steric blockers can also be blocked by recruitment of large protein binding partners. This example is an antibody that binds to serum albumin; The antibody itself may or may not be large enough to block activation or binding by itself, but recruitment of albumin allows sufficient steric blocking.

As used and described herein, a "half-life extending element" refers to, for example, its size (e.g., becoming more than a kidney filtration block), shape, hydrodynamic radius, charge or absorption, biodistribution, metabolism and removal. It is part of a chimeric polypeptide that increases serum half-life and improves pK by changing parameters.

As used herein, the terms “activatable”, “activating”, “inducing” and “inducible” refer to proteins, ie cytokines that are part of the fusion protein, bind to their receptors and cleavage of additional elements from the fusion protein. Refers to the ability to induce activity.

As used herein, a “plasmid” or “viral vector” is an agent comprising a promoter that transfers a disclosed nucleic acid into a cell without degradation and produces expression of the nucleic acid molecule and/or polypeptide in the cell to which it is transferred.

As used herein, the terms "peptide", "polypeptide" or "protein" are used broadly to mean two or more amino acids linked by peptide bonds. Proteins, peptides and polypeptides are also used interchangeably herein to refer to amino acid sequences. It should be appreciated that the term polypeptide is not used herein to suggest a specific size or number of amino acids comprising a molecule, and that the peptides of the invention may contain up to several amino acid residues or more.

As used throughout, “subject” refers to vertebrates, more specifically mammals (eg, humans, horses, cats, dogs, cows, pigs, sheep, goats, mice, rabbits, rats and guinea pigs), birds, It can be reptiles, amphibians, fish and other animals. This term does not refer to a specific age or gender. Thus, it is intended to include adult and neonatal subjects, whether male or female.

As used herein, “patient” or “subject” may be used interchangeably and may refer to a subject having a disease or disorder (eg, cancer). The term patient or subject includes human and veterinary subjects.

As used herein, the terms “treatment”, “treat” or “treating” refer to a method of reducing the effects of a disease or condition or symptoms of a disease or condition. Thus, in the disclosed methods, treatment is at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60, at the severity of the established disease or condition or symptoms of the disease or condition. %, at least about 70%, at least about 80%, at least about 90%, or a substantially complete reduction. For example, a method of treating a disease is considered treatment if there is a 10% reduction in one or more symptoms of the disease in a subject compared to a control. Thus, the reduction is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100% or any decrease between 10% and 100% compared to the native or control level. It can be a percentage. It is understood that treatment does not necessarily mean cure or complete resection of a disease, condition, or symptom of a disease or condition.

As used herein, the terms “prevent”, “preventing” and “prevention” of a disease or disorder refer to an action of inhibiting or delaying the onset or worsening of one or more symptoms of the disease or disorder, eg, Refers to the administration of a chimeric polypeptide or a nucleic acid sequence encoding a chimeric polypeptide, which occurs prior to or approximately simultaneously with a subject beginning to exhibit one or more symptoms of a disease or disorder.

As used herein, reference to “reducing”, “reducing” or “inhibiting” is at least about 10%, at least about 20%, at least about 30%, at least about 40 compared to the appropriate control level. %, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90% or more. These terms may include, but not necessarily, complete elimination of a function or property, such as an agent activity.

"Attenuated cytokine receptor agonists" are cytokine receptor agonists that have reduced receptor agonist activity compared to naturally occurring agonists of cytokine receptors. The attenuated cytokine agonist may have an agonist activity of at least about 10X, at least about 50X, at least about 100X, at least about 250X, at least about 500X, at least about 1000X or less compared to the naturally occurring agonist of the receptor. . When a fusion protein containing a cytokine polypeptide as described herein is described as having “attenuated” or “attenuated activity”, it is meant that the fusion protein is an attenuated cytokine receptor agonist.

An “intact fusion protein” is a fusion protein without domains removed therein, for example by protease cleavage. The domain may be removable by protease cleavage or other enzymatic activity, but this does not occur if the fusion protein is “intact”.

As used herein, “moiety” refers to a portion of a molecule that has distinct functions within a molecule, and that function can be performed by that moiety in the context of another molecule. A moiety may be a chemical entity with a specific function or a part of a biological molecule with a specific function. For example, a “blocking moiety” in a fusion protein is a portion of a fusion protein capable of blocking the activity of some or all of the fusion polypeptide. It can be a protein domain such as serum albumin. Blocking can be accomplished with steric blockers or with specific blockers. Steric blockers are not based on specific binding, but block according to size and location; One example is serum albumin. Certain blocking agents block due to specific interactions with the moiety being blocked. Certain blockers must be tailored to a particular cytokine or active domain; The steric blocker can be used regardless of the payload if it is large enough.

In general, the therapeutic use of cytokines is largely limited by their systemic toxicity. For example, TNF was originally discovered due to its ability to induce hemorrhagic necrosis of some tumors and its in vitro cytotoxic effect on other tumor lineages, but was later demonstrated to have potent pro-inflammatory activity, resulting in an overproduction state. If so, it may have a dangerous effect on the human body. Systemic toxicity is a fundamental problem in the use of pharmacologically active amounts of cytokines in humans, so new derivatives and therapeutic strategies aimed at reducing the toxic effects of this class of biological effects while maintaining their therapeutic efficacy. Is currently being evaluated.

IL-2 exerts both stimulating and regulatory functions in the immune system and, along with other members of the common γ chain (γc) cytokine family, is the center of immune homeostasis. IL-2 is a trimeric receptor consisting of the chains IL-2Rα (CD25), IL-2Rβ (CD122) and IL-2Rγ (γc, CD132) or an IL-2 receptor consisting of dimer βγ IL-2R (1, 3) It mediates its action by binding to (IL-2R). Both IL-2R variants are capable of transmitting signals upon binding to IL-2. However, trimer αβγ IL-2R has about 10-100 times higher affinity for IL-2 than dimer βγ IL-2R (3), which indicates that CD25 binds its high affinity to the receptor of IL-2. But is not important for signal transduction. Trimeric IL-2R is found on activated T cells and on CD4+ folkhead box P3 (FoxP3)+ T regulatory cells (Treg), which are sensitive to IL-2 in vitro and in vivo. Conversely, antigen-experience (memory) CD8+, CD44 hypermembrane-phenotype (MP) CD8+, and natural killer (NK) cells are endowed with high levels of the dimer βγ IL-2R, and these cells are also conferred in vitro and in vivo . It reacts violently to IL-2.

The expression of high affinity IL-2R is important to allow T cells to respond to low concentrations of IL-2 that are temporarily available in vivo. IL-2Rα expression is absent in naive and memory T cells, but is induced after antigen activation. IL-2Rβ is constitutively expressed by NK, NKT and memory CD8+ T cells, but is also induced on naive T cells after antigen activation. γc is much less tightly regulated and constitutively expressed by all lymphocyte cells. When high affinity IL-2R is induced by an antigen, IL-2R signaling upregulates the expression of IL-2Rα in part through Stat5-dependent regulation of Il2ra transcription (Kim et al., 2001). This process represents a mechanism for maintaining the expression of high affinity IL-2R and maintaining IL-2 signaling while the source of IL-2 remains.

IL-2 is captured by IL-2Rα through a large hydrophobic binding surface surrounded by the periphery of the pole, resulting in a relatively weak interaction (Kd 10-8 M) with fast on-off binding kinetics. However, the IL-2Rα-IL-2 binary complex results in a very small conformational change in IL-2 that enhances the association with IL-2Rβ through a distinct polar interaction between IL-2 and IL-2Rβ. The quasi-high affinity of the IL2/α/β trimer complex (i.e., Kd ~ 300 pM) was determined by Ciardelli's data that the trimer complex was either α chain alone (Kd = 10 nM) or β chain alone (Kd = 450 nM). It clearly indicates that it is more stable than either IL2 bound to. In any case, the IL2/α/β trimer then recruits the γ chain into a quaternary complex capable of signaling, which is facilitated by a large complex binding site on the IL2-binding β chain to the γ chain.

등, 1995; Yu 및 Malek, 2001). 따라서, 지속적인 IL-2R 신호전달을 필요로 하는 이들 기능적 활성은 IL-2Rα와 계합하고 추가적인 IL-2-IL-2R 신호전달 복합체를 형성하기 위해 계속된 IL-2의 공급원을 필요로 한다.When reduced, the ternary IL-2Rα-IL-2Rβ-IL-2 complex then mobilizes γc through weak interactions with IL-2 and strong interactions with IL-2Rβ, resulting in stable fourth-order high-affinity IL-2R ( 10pM Kd 10-11M) is produced. The formation of the high affinity quaternary IL-2-IL-2R complex induces signaling through the tyrosine kinases Jak1 and Jak3, respectively, associated with IL-2Rβ and γc (Nelson and Willerford, 1998). The quaternary IL-2-IL-2R complex is rapidly internalized, resulting in rapid degradation of IL-2, IL-2Rβ and γc, but IL-2Rα is recycled to the cell surface (

Et al., 1995; Yu and Malek, 2001). Thus, these functional activities that require sustained IL-2R signaling require a continued source of IL-2 to engage IL-2Rα and form additional IL-2-IL-2R signaling complexes.

Regulatory T cells actively inhibit the activation of the immune system and prevent pathological autoreactions and subsequent autoimmune diseases. Development of drugs and methods for selectively activating regulatory T cells for the treatment of autoimmune diseases is the subject of intensive research, and the development of the present invention capable of selectively delivering active interleukins to inflammatory sites has mainly failed. Regulatory T cells (Treg) are a class of CD4+CD25+ T cells that inhibit the activity of other immune cells. Tregs are the center of immune system homeostasis and play an important role in maintaining resistance to self-antigens and regulating immune responses to foreign antigens. Several autoimmune and inflammatory diseases, including type 1 diabetes (T1D), systemic lupus erythematosus (SLE), and graft-versus-host disease (GVHD), have been shown to lack Treg cell count or Treg function.

As a result, there is great interest in the development of therapeutics that improve the number and/or function of Treg cells. One therapeutic approach for the autoimmune disease under investigation is the transplantation of autologous tissue, ex vivo-expanded Treg cells (Tang, Q., et al., 2013, Cold Spring Harb. Perspect. Med., 3:1-15 ). While this approach treats animal models of the disease and has been shown to be promising in several early stage human clinical trials, it requires personalized treatment with the patient's own T cells and is invasive and technically complex. Another approach is treatment with low-dose interleukin-2 (IL-2). Treg cells characteristically express a high constitutive level of IL2Rαβγ, a high affinity IL-2 receptor consisting of subunits IL2Rα (CD25), IL2Rβ (CD122) and IL2Rγ (CD132), and Treg cell growth is IL-2 dependent. Appeared (Malek, TR, et al., 2010, Immunity, 33:153-65).

Conversely, immune activation was also achieved using IL-2, and recombinant IL-2 (Proleukin®) has been approved for the treatment of certain cancers. High-dose IL-2 is used in the treatment of patients with metastatic melanoma and metastatic renal cell carcinoma with a long-term effect on overall survival.

Chronic GVHD (Koreth, J., et al., 2011, N Engl J Med., 365:2055-66) and HCV-associated autoimmune vasculitis patients (Saadoun, D., et al., 2011, N Engl J Med., 365: 2067-77), a clinical trial of low-dose IL-2 treatment demonstrated increased Treg levels and signs of clinical efficacy. A new clinical trial has begun examining the efficacy of IL-2 in a number of other autoimmune and inflammatory diseases. The rationale for the use of the so-called low-dose IL-2 utilizes the high IL-2 affinity of the trimeric IL-2 receptor constitutively expressed on Tregs, leaving other T cells that do not express the high affinity receptor in the inactive state. Was to do. Aldesleukin, the recombinant form of IL-2 used in these tests (sold as Proleukin® by Prometheus Laboratories, San Diego, Calif.) is associated with high toxicity. Aldesleukin is approved for the treatment of metastatic melanoma and metastatic kidney cancer, but its side effects are so severe that its use is only recommended in hospitals with intensive care units (web address: www.proleukin.com/assets/pdf/proleukin.pdf) .

Clinical trials of IL-2 in autoimmune diseases have shown that Treg cells respond to lower concentrations of IL-2 than many other immune cell types due to their expression of IL2R alpha. A lower dose was used (Klatzmann D, 2015 Nat Rev Immunol. 15:283-94). However, even these low doses resulted in safety and tolerability issues, and the treatment used was a daily subcutaneous injection in a chronic or intermittent 5-day course of treatment. Thus, there is a need for enhancing Treg cell number and function, targeting Treg cells more specifically than IL-2, and treating autoimmune diseases that are safer, more resistant and less frequently administered.

One approach proposed to improve the therapeutic index of IL-2-based therapy for autoimmune diseases is to use variants of IL-2 that are selective for Treg cells over other immune cells. The IL-2 receptor is expressed on a variety of different immune cell types, including T cells, NK cells, eosinophils and monocytes, and this broad expression pattern is likely to contribute to multiple effects on the immune system and high systemic toxicity. In particular, activated T effector cells express IL2Rαβγ like lung epithelial cells. However, activating T effector cells is directly contrary to the goal of down-regulating and controlling the immune response, and activating pulmonary epithelial cells results in known dose-limiting side effects of IL-2, including pulmonary edema. In fact, the major side effect of high-dose IL-2 immunotherapy is vascular leakage syndrome (VLS), which accumulates intravascular fluid in organs such as the lungs and liver, resulting in subsequent lung swelling and liver cell damage. There is no treatment for VLS other than recovery of IL-2. Low-dose IL-2 therapy was tested in patients to avoid VLS, but sacrificed suboptimal treatment results.

According to the literature, VLS is believed to be due to the release of proinflammatory cytokines from IL-2-activated NK cells. However, there is strong evidence that pulmonary edema results from direct binding of IL-2 to lung endothelial cells expressing low to moderate levels of functional αβγ IL-2R. In addition, pulmonary edema associated with the interaction of IL-2 with pulmonary endothelial cells can block binding to CD25 in CD25-deficient host mice with anti-CD25 monoclonal antibody (mAb) or CD122-specific IL-2/anti- It was eliminated by the use of the IL-2 mAb (IL-2/mAb) complex, thus preventing VLS.

Treatment with interleukin cytokines other than IL-2 was more limited. IL-15 exhibits similar immune cell stimulating activity to that of IL-2, but does not have the same inhibitory effect, making it a promising immunotherapy candidate. Clinical trials of recombinant human IL-15 for the treatment of metastatic malignant melanoma or renal cell carcinoma have demonstrated noticeable changes in immune cell distribution, proliferation, and activation and suggested potential anti-tumor activity (Conlon et al., 2014). . IL-15 is currently in clinical trials to treat various types of cancer. However, IL-15 therapy is known to be associated with unwanted toxic effects such as exacerbating certain leukemias, graft-versus-host disease, hypotension, thrombocytopenia and liver damage. (Mishra A., et al., Cance Cell, 2012, 22(5):645-55; Alpdogan O. et al., Blood, 2005, 105(2):866-73; Conlon KC et al., J Clin Oncol, 2015, 33 (1):74-82.)

The direct use of IL-2 as an agent that binds to IL-2R and modulates the immune response therapeutically has been problematic due to well-proven therapeutic risks such as, for example, its short serum half-life and high toxicity. These risks also limited the therapeutic development and use of other cytokines. New types of cytokines are needed to reduce these risks. Compositions and methods comprising IL-2 and IL-15 and other cytokines, functional fragments and muteins of cytokines, as well as conditionally active cytokines designed to address these risks and provide the necessary immunomodulatory therapeutics are disclosed herein It is disclosed in

The present invention includes, for example, cytokine blocking moieties, such as steric blocking polypeptides, serum half-life extending polypeptides, targeting polypeptides, protease cleavable linkers using direct IL-2 therapy and other cytokines. It is designed to address the shortcomings of therapies using linking polypeptides, and combinations thereof. Interleukin (e.g., IL-2, IL-7, IL-12, IL-15, IL-18, IL-21 IL-23), interferon (IFN including IFNalpha, IFNbeta and IFNgamma), tumor Necrosis factor (e.g., TNFalpha, lymph toxin), transforming growth factor (e.g., TGFbeta1, TGFbeta2, TGFbeta3), chemokines (CXC motif chemokine 10 (CXCL10), CCL19, CCL20, CCL21) and granulocyte macrophage-colony stimulating factor (GM-CS) are very potent when administered to patients. As used herein, “chemokine” refers to a family of small cytokines that have the ability to induce directed chemotaxis in nearby reactive cells. Cytokines can provide potent treatments, but they are difficult to control clinically and have unwanted effects that limit the clinical use of cytokines. The present disclosure relates to a new type of cytokine that can be used in patients with reduced or eliminated undesired effects. In particular, the present disclosure relates to a cytokine having a reduced cytokine receptor activation activity compared to the corresponding cytokine or a chimeric polypeptide containing an active fragment or mutein of a cytokine (fusion protein), a nucleic acid encoding a fusion protein And it relates to a pharmaceutical composition comprising a pharmaceutical formulation of the above. However, under selected conditions or in a selected biological environment, chimeric polypeptides often activate their cognate receptors with the same or higher potency as the corresponding naturally occurring cytokines. As described herein, it is typically not under selected conditions, such as present at a desired site of cytokine activity (e.g., an inflammation site or tumor), but under normal conditions, of a cytokine, an active fragment thereof, or a mutein. It is achieved using cytokine blocking moieties that block or inhibit receptor activating functions.

Chimeric polypeptides and nucleic acids encoding chimeric polypeptides can be prepared using any suitable method. For example, nucleic acids encoding chimeric polypeptides can be prepared using recombinant DNA techniques, synthetic chemistry, or a combination of these techniques, and can be expressed in a suitable expression system such as in CHO cells. Chimeric polypeptides can be similarly prepared by expression of suitable nucleic acids, for example using synthetic or semi-synthetic chemistry techniques and the like. In some embodiments, the blocking moiety can be attached to the cytokine polypeptide through a sortase-mediated conjugation. "Sortase" is a transpeptidase that modifies a protein by recognizing and cleaving a carboxyl-terminal classification signal embedded in or attached to a terminal protein or peptide. Sortase A handles the cleavage of the LPXTG motif (SEQ ID NO: 125) (where X is any standard amino acid) between the Thr and Gly residues on the target protein with the transient attachment of the Thr residue to the active site Cys residue on the enzyme. Catalyzes the formation of the enzyme-thioacyl intermediate. In order to complete the transpeptization and generate a peptide-monomer conjugate, a biomolecule with an N-terminal nucleophilic group, typically an oligoglycine motif, attacks the intermediate to displace Sortase A and binds the two molecules.

To form a cytokine-blocking moiety fusion protein, the cytokine polypeptide is first tagged at the N-terminus with the polyglycine sequence, or alternatively at the C-terminus with the LPXTG motif (SEQ ID NO: 125). A blocking moiety or other element is attached to each peptide that acts as an acceptor site for the tagged polypeptide. For conjugation to the domain carrying the LPXTG acceptor peptide (SEQ ID NO: 125) attached via the N-terminus, the polypeptide will be tagged with an N-terminal poly-glycine stretch. For conjugation to a domain carrying a poly-glycine peptide attached via the C-terminus, the polypeptide will be tagged at its C-terminus with the LPXTG sortase recognition sequence (SEQ ID NO: 125). Sortases that recognize poly-glycine and LPXTG (SEQ ID NO: 125) sequences will form a peptide bond between the polymer-peptide and the tagged polypeptide. The sortase reaction occurs at room temperature after cleaving the glycine residue as an intermediate.

A variety of mechanisms can be used to eliminate or reduce the inhibition caused by the blocking moiety. For example, the pharmaceutical composition comprises an IL-2 polypeptide and a blocking moiety, e.g., a protease cleavage site located between the IL-2 polypeptide and the IL-2 blocking moiety or within the IL-2 blocking moiety. It may comprise a steric blocking moiety with a linker capable of protease cleavage. When the protease cleavage site is cleaved, the blocking moiety can dissociate from the cytokine, and the cytokine can then activate the cytokine receptor. Cytokine moieties can also be blocked by certain blocking moieties, such as antibodies that bind to epitopes found on related cytokines.

Any suitable linker can be used. For example, the linker may be a glycine-glycine, a sortase-recognition motif, or a sortase-recognition motif and a peptide sequence (Gly ₄ Ser) _n (SEQ ID NO: 126) or (Gly ₃ Ser) _n (SEQ ID NO: 127). May include, wherein n is 1, 2, 3, 4 or 5. Typically, the sortase-recognition motif comprises the peptide sequence LPXTG (SEQ ID NO: 125), where X is any amino acid. In some embodiments, the covalent bond is between a reactive lysine moiety attached to the C-terminus of the cytokine polypeptide and a reactive aspartic acid attached to the N-terminus of a blocking agent or other domain. In another embodiment, the covalent bond is between a reactive aspartic acid residue attached to the N-terminus of the cytokine polypeptide and a reactive lysine residue attached to the C-terminus of the blocker or other domain.

Thus, as detailed herein, the cytokine blocking moiety (eg, IL-2 blocking moiety) used may be a steric blocker. As used herein, a “stereoblocker” may be covalently linked to a cytokine polypeptide, either directly or indirectly through another moiety such as a linker, for example in the form of a chimeric polypeptide (fusion protein), However, otherwise it refers to a polypeptide or a polypeptide moiety that does not covalently bind to a cytokine polypeptide. Steric blockers can non-covalently bind to cytokine polypeptides, for example via electrostatic, hydrophobic, ionic or hydrogen bonding. Steric blockers typically inhibit or block the activity of cytokine moieties due to their proximity and relative size to the cytokine moiety. Steric inhibition of cytokine moieties spatially separates cytokine moieties from steric blockers, such as by enzymatically cleaving the fusion protein containing the steric blocker and the cytokine polypeptide at the site between the steric blocker and the cytokine polypeptide. It can be removed by separating.

As described in more detail herein, the blocking function may be combined with or due to the presence of additional functional components in the pharmaceutical composition such as targeting domains, serum half-life extending elements and protease-cleavable linking polypeptides. . For example, serum half-life extending polypeptides can also be steric blockers.

Various factors ensure the delivery and activity of IL-2 preferentially at the desired site of IL-2 activity and severely limit systemic exposure to interleukins through blocking and/or targeting strategies linked preferentially to serum half-life extension strategies. In this serum half-life extension strategy, the blocked version of the interleukin circulates for an extended period of time (preferably at least 1-2 weeks) while the activated version has a typical serum half-life of interleukin.

Serum half-life Compared to the extended version, the serum half-life of IL-2 administered intravenously is only -10 minutes due to its distribution to the entire body extracellular space, mainly -15 L in average-sized adults. Subsequently, IL-2 is metabolized by the kidneys with a half-life of -2.5 hours. (Smith, K. "Interleukin 2 immunotherapy." Therapeutic Immunology 240 (2001)). By another measure, IL-2 has a very short plasma half-life of 85 minutes for intravenous administration and 3.3 hours of subcutaneous administration (Kirchner, GI, et al., 1998, Br J Clin Pharmacol. 46:5-10). In some embodiments of the present invention, the half-life extending element is linked to the interleukin via a linker that is cleaved at the site of action (e.g., by inflammation-specific or tumor-specific proteases) to enable full activity of the interleukin at the desired site. It releases and also separates it from the half-life extension of the uncut version. In this embodiment, the fully active and free interleukin will have very different pharmacokinetic (pK) properties-half-life of hours, not weeks. In addition, exposure to active cytokines is limited to sites of desired cytokine activity (eg, sites of inflammation or tumors) and systemic exposure to active cytokines, and associated toxicity and side effects are reduced.

Other cytokines envisioned in the present invention are pharmacology similar to IL-2 (e.g., Blood 2011 117:4787-4795; doi: IL-15 reported by doi.org/10.1182/blood-2010-10-311456) ) And therefore, the design of the present invention overcomes the drawbacks of using these agents directly, may have an extended half-life and/or be targeted to the site of desired activity (e.g., site of inflammation or site of tumor). It provides a possible chimeric polypeptide.

If desired, IL-2 is generally directed to an IL-2R complex or specifically to one of the three IL-2R subunits with an affinity different from that of the corresponding wild-type IL-2, for example to selectively activate Tregs or Teff. Can be manipulated to combine. For example, IL-2 polypeptides mentioned as having a higher affinity for the trimeric form of the Il-2 receptor compared to the dimeric beta/gamma form of the IL-2 receptor compared to wild-type IL-2 have sequence Number: 1 (a mature IL-2 protein comprising amino acids 21-153 of human IL-2 with Uniprot accession number P60568-1) may have an amino acid comprising one of the following mutation sets: (a ) K64R, V69A, and Q74P; (b) V69A, Q74P, and T101A; (c) V69A, Q74P, and I128T; (d) N30D, V69A, Q74P, and F103S; (e) K49E, V69A, A73V, and K76E; (f) V69A, Q74P, T101A, and T133N; (g) N30S, V69A, Q74P, and I128A; (h) V69A, Q74P, N88D, and S99P; (i) N30S, V69A, Q74P, and I128T; (j) K9T, Q11R, K35R, V69A, and Q74P; (k) A1T, M46L, K49R, E61D, V69A, and H79R; (l) K48E, E68D, N71T, N90H, F103S, and I114V; (m) S4P, T10A, Q11R, V69A, Q74P, N88D, and T133A; (n) E15K, N30S Y31H, K35R, K48E, V69A, Q74P, and I92T; (o) N30S, E68D, V69A, N71A, Q74P, S75P, K76R, and N90H; (p) N30S, Y31C, T37A, V69A, A73V, Q74P, H79R, and I128T; (q) N26D, N29S, N30S, K54R, E67G, V69A, Q74P, and I92T; (r) K8R, Q13R, N26D, N30T, K35R, T37R, V69A, Q74P, and I92T; And (s) N29S, Y31H, K35R, T37A, K48E, V69A, N71R, Q74P, N88D, and I89V. This approach includes interleukins (e.g., IL-2, IL-7, IL-12, IL-15, IL-18, IL-23), interferons (IFN including IFNalpha, IFNbeta and IFNgamma), tumor Necrosis factor (e.g., TNFalpha, lymphotoxin), transforming growth factor (e.g., TGFbeta1, TGFbeta2, TGFbeta3), and granulocyte macrophage-colonial stimulating factor (GM-CS). It can also be applied to make muteins of other cytokines. For example, muteins with a desired binding affinity for cognate receptors can be prepared.

As mentioned above, any mutant IL-2 polypeptide disclosed herein may comprise the described sequence; They may also be limited to the sequence described or otherwise identical to SEQ ID NO: 1. Moreover, any of the mutant IL-2 polypeptides disclosed herein may optionally comprise a substitution of a cysteine residue at position 125 for another residue (e.g., serine) and/or optionally at position 1 of SEQ ID NO: 1 May include a deletion of an alanine residue in.

Another approach to improving the therapeutic index of IL-2 based therapy is to optimize the pharmacokinetics of the molecule to maximize activation of Treg cells. Early studies of IL-2 action demonstrated that IL-2 stimulation of human T cell proliferation in vitro required exposure to an effective concentration of IL-2 for at least 5-6 hours (Cantrell, DA, et al., 1984, Science, 224: 1312-1316). When administered to human patients, IL-2 has a very short plasma half-life of 85 minutes for intravenous administration and 3.3 hours for subcutaneous administration (Kirchner, GI, et al., 1998, Br J Clin Pharmacol. 46:5-10). . Because of its short half-life, maintaining circulating IL-2 above the levels necessary to stimulate T cell proliferation for the required period of time results in peak IL-2 levels much higher than the EC50 for Treg cells or requires frequent administration. It requires a high dose to do. These high IL-2 peak levels may activate the IL2Rβγ receptor and may have other unintended or adverse effects, such as VLS as mentioned above. IL-2 analogs or multifunctional proteins with IL-2 attached to a domain capable of binding to the FcRn receptor, with a longer circulating half-life than IL-2, at lower doses and at lower peak levels than IL-2. Target drug concentration can be achieved over a specific period of time. Thus, these IL-2 analogs will require lower doses or less frequent administration than IL-2 to effectively stimulate Treg cells. Less frequent subcutaneous administration of IL-2 drugs may also be more resistant to patients. Therapies with these properties will translate into clinically improved pharmacological efficacy, reduced toxicity, and improved patient compliance to treatment. Alternatively, IL-2 or muteins of IL-2 (hereinafter "IL-2 ^* ") can be selectively targeted to an intended site of action (eg, site of inflammation). This targeting can be achieved by one of several strategies involving the addition of a domain to an administered agent comprising a blocker of cleaved IL-2 (or mutein), or by a targeting domain or a combination of the two.

In some embodiments, the IL-2 ^* partial agonist can be tailored to bind with a higher or lower affinity depending on the desired target; For example, IL-2 ^* can be engineered to bind with enhanced affinity to one of the receptor subunits other than the other receptor. Unlike full agonists or full antagonists, these types of partial agonists provide the ability to adjust signal properties to a magnitude that elicits the desired functional properties without meeting the threshold for the undesired properties. Given the differential activity of partial agonists, the repertoire of IL-2 variants can be engineered to exhibit unique signaling activity at even finer levels ranging from near full to partial to full antagonism.

In some embodiments, IL-2 ^* has an altered affinity for IL-2Rα. In some embodiments, IL-2 ^* has a higher affinity for IL-2Rα than wild type IL-2. In another embodiment, IL-2 ^* has an altered affinity for IL-2Rβ. In one embodiment, IL-2 ^* has enhanced binding affinity to the N-terminus of IL-2Rβ, eg, IL-2Rβ, which eliminates the functional requirement for IL-2Rα. In yet another embodiment, has the increased binding affinity for the IL-2Rβ shows a reduced binding to the IL-2Rγ, whereby the IL-2Rβγ heterodimer dimer Chemistry and IL-2 may be defective in signal transduction ^- a Is created.

Blocking moieties described in more detail below can also be used to favor binding to or activation of one or more receptors. In one embodiment, a blocking moiety is added such that IL-2Rβγ binding or activation is blocked but IL-2Rα binding or activation is not altered. In another embodiment, a blocking moiety is added such that IL-2Rα binding or activation is reduced. In another embodiment, a blocking moiety is added such that binding to and/or activation of all three receptors is inhibited. This blocking can be mitigated in certain circumstances by removal of the blocking moiety, for example by proteolytic cleavage of a linker that connects one or more blocking moieties to cytokines.

A similar approach can be applied to improve other cytokines, especially for use as immunostimulants, eg to treat cancer. For example, in this embodiment, cytokines (e.g., IL-2, IL-7, IL-12, IL-15, IL-18, IL-21 IL-23, IFN alpha, IFN beta and IFN gamma , TNFalpha, lymphotoxin, TGFbeta1, TGFbeta2, TGFbeta3 GM-CSF, CXCL10, CCL19, CCL20 and CCL21 pharmacokinetics and/or pharmacodynamics at the desired active site, such as in tumors, but preferably systemic It can be tailored to maximally activate effector cells (eg, effector T cells, NK cells) and/or cytotoxic immune response enhancing cells (eg, inducing dendritic cell maturation) at sites other than those.

Thus, at least one cytokine polypeptide, such as an interleukin (e.g., IL-2, IL-7, IL-12, IL-15, IL-18, IL-21, IL-23), interferon (IFNalpha , IFN including IFNbeta and IFNgamma), tumor necrosis factor (e.g., TNFalpha, lymphotoxin), transforming growth factor (e.g., TGFbeta1, TGFbeta2, TGFbeta3), chemokines ( Pharmaceutical compositions comprising, for example, CXCL10, CCL19, CCL20, CCL21) and granulocyte macrophage-colony stimulating factor (GM-CS) or functional fragments or muteins of the foregoing are provided herein. Polypeptides also typically comprise at least one linker amino acid sequence, wherein the amino acid sequence can in certain embodiments be cleaved by an endogenous protease. In one embodiment, the linker is an amino acid comprising HSSKLQ (SEQ ID NO: 25), GPLGVRG (SEQ ID NO: 128), IPVSLRSG (SEQ ID NO: 129), VPLSLYSG (SEQ ID NO: 130), or SGESPAYYTA (SEQ ID NO: 131). Include sequence. In other embodiments, the chimeric polypeptide further contains a blocking moiety, eg, a steric blocking polypeptide moiety capable of blocking the activity of an interleukin polypeptide. For example, the blocking moiety can comprise a human serum albumin (HSA) binding domain or optionally branched or multi-armed polyethylene glycol (PEG). Alternatively, the pharmaceutical composition comprises a first cytokine polypeptide or fragment thereof, and a blocking moiety, e.g., a steric blocking polypeptide moiety, wherein the blocking moiety is a cytokine for a cytokine receptor. Blocks the activity of the polypeptide, and in certain embodiments the blocking moiety comprises a protease cleavable domain. In some embodiments, blocking and reducing cytokine activity is achieved simply by attaching an additional domain with a very short linker to the N or C terminus of the interleukin domain. In this embodiment, the blocking agent is expected to be mitigated by protease digestion of the blocking moiety or short linker that binds the blocking agent to the interleukin. When a domain is truncated or released, it can no longer achieve blocking of cytokine activity.

Pharmaceutical compositions, for example chimeric polypeptides, may comprise two or more cytokines, which may be the same cytokine polypeptide or different cytokine polypeptides. For example, two or more different types of cytokines have complementary functions. In some examples, the first cytokine is IL-2 and the second cytokine is IL-12. In some embodiments, each of the two or more different types of cytokine polypeptides has an activity that modulates the activity of the other cytokine polypeptide. In some examples of chimeric polypeptides containing two cytokine polypeptides, the first cytokine polypeptide is T-cell activation and the second cytokine polypeptide is non-T-cell-activation. In some examples of chimeric polypeptides containing two cytokine polypeptides, the first cytokine is a chemoattractant, such as CXCL10 and the second cytokine is an immune cell activator.

Preferably, cytokine polypeptides (including functional fragments) contained in the fusion proteins disclosed herein are not mutated or engineered to alter the properties of naturally occurring cytokines, including receptor binding affinity and specificity or serum half-life. Does not. However, for example, changes in the amino acid sequence from naturally occurring (including wild-type) cytokines may be allowed to facilitate cloning and achieve the desired level of expression.

CD25 binding

CD25 binding is often not recommended in modified IL-2 constructs. In contrast, the IL-2 polypeptides described herein are preferably unmodified to avoid CD25 binding. Preferably, the IL-2 polypeptide described herein binds to CD25. Typically, the IL-2 fusion proteins described herein are capable of CD25 binding and blocking is for interactions with IL-2R beta and gamma (CD122 and CD132).

Blocking moiety

The blocking moiety can be any moiety that inhibits the ability of a cytokine to bind and/or activate its receptor. A blocking moiety can inhibit the ability of a cytokine to bind and/or activate its receptor by sterically blocking and/or non-covalently binding to a cytokine. Examples of suitable blocking moieties include full-length or cytokine-binding fragments or muteins of cognate receptors of cytokines. Polyclonal antibodies that bind cytokines, recombinant antibodies, human antibodies, humanized antibodies, single chain variable fragments (scFv), single-domain antibodies such as heavy chain variable domain (VH), light chain variable domain (VL) and camel-type nano Antibodies and fragments thereof comprising the body's variable domain (VHH), dAb, and the like can also be used. Other suitable antigen-binding domains that bind cytokines can also be used, and non-immunoglobulin proteins that mimic antibody binding and/or structure, such as anticalin, apilin, apibody molecule, apimer, afitin, Alphabody, Avimer, DARPin, Finomer, Kunitz domain peptide, monobody and binding domains based on other engineered scaffolds such as SpA, GroEL, fibronectin, lipocalin and CTLA4 scaffold. Further examples of suitable blocking polypeptides include polypeptides that sterically inhibit or block binding of cytokines to their cognate receptors. Advantageously, this moiety can also function as a half-life extending element. For example, a peptide modified by conjugation to a water-soluble polymer such as PEG can sterically inhibit or prevent the binding of cytokines to their receptors. Polypeptides or fragments thereof having a long serum half-life, such as serum albumin (human serum albumin), immunoglobulin Fc, transferrin, etc., as well as fragments and muteins of such polypeptides can also be used. For example, antibodies and antigen-binding domains that bind to proteins with long serum half-lives such as HSA, immunoglobulins or transferrin, or receptors that are recycled to the plasma membrane such as FcRn or transferrin receptors, are also cytokines, especially when binding to their antigens. Can be suppressed. Examples of such antigen-binding polypeptides include single chain variable fragments (scFv), single-domain antibodies such as heavy chain variable domain (VH), light chain variable domain (VL) and variable domain of camel-type nanobody (VHH), dAb, etc. Includes. Other suitable antigen-binding domains that bind cytokines can also be used, and non-immunoglobulin proteins that mimic antibody binding and/or structure, such as anticalin, apilin, apibody molecule, apimer, afitin, Alphabody, abimer, DARPin, finomer, Kunitz domain peptide, monobody and binding domains based on other engineered scaffolds such as SpA, GroEL, fibronectin, lipocalin and CTLA4 scaffold.

In an illustrative example, when IL-2 is a cytokine in the chimeric polypeptide, the blocking moiety is the alpha chain of the IL-2 receptor (IL-2Rα) or the beta of the IL-2 receptor (IL-2Rβ) or the gamma chain ( IL-2Rγ), anti-IL-2 single-domain antibody (dAb) or scFv, Fab, anti-CD25 antibody or fragment thereof, and full length or fragments or muteins such as anti-HAS dAb or scFv.

Further aspects of the invention

1.A fusion protein comprising a cytokine moiety operably linked to a binding moiety, wherein the binding moiety comprises a non-CDR loop and a cleavable linker, wherein the binding moiety carries a cytokine to its receptor. A fusion protein capable of masking binding and/or activation of receptors by cytokines.

2. The fusion protein of aspect 1, wherein the binding moiety is a natural peptide, a synthetic peptide, an engineered scaffold or an engineered bulk serum protein.

3. According to aspect 1 or 2, the engineered scaffold is sdAb, scFv, Fab, VHH, fibronectin type III domain, immunoglobulin-like scaffold, DARPin, cystine knot peptide, lipocalin, 3-helix bundle scaffold , A protein G-related albumin-binding module, or a DNA or RNA aptamer scaffold.

4. The fusion protein according to any one of aspects 1-2, wherein the binding moiety is capable of binding to a bulk serum protein.

5. The fusion protein of any one of aspects 1-3, wherein the non-CDR loop is derived from a variable domain, constant domain, C1-set domain, C2-set domain, I-domain, or any combination thereof.

6. The fusion protein according to any one of aspects 1-4, wherein the binding moiety further comprises a complementarity determining region (CDR).

7. The fusion protein according to aspect 5, wherein the binding moiety is capable of binding to a bulk serum protein.

8. The fusion protein according to aspect 6, wherein the bulk serum protein is a half-life extending protein.

9. The fusion protein according to aspect 6 or 7, wherein the bulk serum protein is albumin, transferrin, factor XIII or fibrinogen.

10. The fusion protein of any one of embodiments 5-8, wherein the CDR loops provide a binding site specific for a bulk serum protein or immunoglobulin light chain, or any combination thereof.

11. The fusion protein according to any one of embodiments 1-9, wherein the cleavable linker comprises a cleavage site.

12. The fusion protein according to aspect 10, wherein the cleavage site is recognized by a protease.

13. The fusion protein of aspect 11, wherein the binding moiety is bound to a cytokine.

14. The fusion protein of embodiment 11 or 11, wherein the binding moiety is covalently linked to a cytokine.

15. The fusion protein according to aspect 11, 11 or 14, wherein the binding moiety is capable of masking binding of a cytokine to its target through specific intermolecular interactions between the binding moiety and the cytokine.

16. The fusion protein of any of embodiments 11-14, wherein the non-CDR loop provides a binding site specific for binding of the moiety to a cytokine.

17. The fusion protein of any one of embodiments 11-15, wherein upon cleavage of the cleavable linker, the binding moiety is separated from the cytokine and the cytokine binds its target.

18. The fusion protein according to any one of aspects 1-16, wherein the cytokine binds to a cytokine receptor.

19. The fusion protein according to aspect 17, wherein the cytokine receptor comprises a type I cytokine receptor, a type I IL receptor, a type II IL receptor, a chemokine receptor, or a tumor necrosis receptor superfamily receptor.

20. The fusion protein of any of embodiments 1-18, wherein the cleavable linker comprises a cleavage site.

21. The fusion protein of aspect 20, wherein the cleavage site is recognized by a protease.

22. The fusion protein according to aspect 21, wherein the protease cleavage site is recognized by serine protease, cysteine protease, aspartate protease, threonine protease, glutamic acid protease, metalloproteinase, gelatinase or asparagine peptide lyase.

23. In aspect 21, the protease cleavage site is cathepsin B, cathepsin C, cathepsin D, cathepsin E, cathepsin K, cathepsin L, kallikrein, hK1, hK10, hK15, plasmin, collagena Type IV collagenase, stromelysin, factor Xa, chymotrypsin-like protease, trypsin-like protease, elastase-like protease, subtilisin-like protease, actinidin, bromelain, calpine , Caspase, caspase-3, Mir1-CP, papain, HIV-1 protease, HSV protease, CMV protease, chymosin, renin, pepsin, matriptase, legumine, plasmexin, nepenthesin, metalloexo Peptidase, metalloendopeptidase, matrix metalloprotease (MMP), MMP1, MMP2, MMP3, MMP8, MMP9, MMP10, MMP11, MMP12, MMP13, MMP14, ADAM10, ADAM17, ADAM12, urokinase plasminogen activator (uPA) , Enterokinase, prostate-specific target (PSA, hK3), interleukin-1β converting enzyme, thrombin, FAP (FAP-α), dipeptidyl peptidase, or dipeptidyl peptidase IV (DPPIV/CD26), II Type transmembrane serine protease (TTSP), neutrophil elastase, cathepsin G, proteinase 3, neutrophil serine protease 4, mast cell chimase, mast cell tryptase, dipeptidyl peptidase and dipeptidyl peptidase IV (DPPIV/ CD26).

24. A conditionally active binding protein comprising a binding moiety (M) comprising a non-CDR loop, a cytokine and a cleavable linker (L), wherein the non-CDR loop is capable of binding a cytokine and , The binding moiety is a conditionally active binding protein capable of inhibiting the binding of a cytokine to its receptor and/or inhibiting the activation of a receptor by a cytokine.

25. The binding protein of aspect 24, wherein the binding moiety is capable of binding to a half-life extending protein.

26. The conditionally active binding protein of aspect 24 or 25, wherein the binding moiety is a natural peptide, a synthetic peptide, an engineered scaffold or an engineered serum bulk protein.

27. The method of embodiment 26, wherein the engineered scaffold is sdAb, scFv, Fab, VHH, fibronectin type III domain, immunoglobulin-like scaffold, DARPin, cystine knot peptide, lipocalin, 3-helix bundle scaffold, protein. A binding protein that is conditionally active, comprising a G-related albumin-binding module or DNA or RNA aptamer scaffold.

28. The conditional of any one of embodiments 24-27, wherein the non-CDR-loop is from a variable domain, constant domain, C1-set domain, C2-set domain, I-domain, or any combination thereof. Binding protein active as.

29. The binding protein of any one of embodiments 24-28, wherein the binding moiety further comprises a complementarity determining region (CDR).

30. The binding protein of any of embodiments 24-29, wherein the binding moiety comprises a binding site specific for bulk serum protein.

31. A conditionally active binding protein according to aspect 30, wherein the bulk serum protein is albumin, transferrin, factor XIII or fibrinogen.

32. The binding protein of any one of embodiments 29-31, wherein the CDRs provide a binding site specific for a bulk serum protein or immunoglobulin light chain, or any combination thereof.

33. The method of any of embodiments 29-32, wherein the binding moiety is conditionally active, capable of masking the binding of the cytokine to its target through specific intermolecular interactions between the binding moiety and the cytokine. Binding protein.

34. The conditionally active binding protein of any of embodiments 29-33, wherein the non-CDR loop provides a binding site specific for binding of the binding moiety to a cytokine.

35. A conditionally active binding protein according to any of embodiments 24-34, wherein the cytokine binds to a cytokine receptor.

36. The binding protein according to aspect 35, wherein the cytokine receptor comprises a type I cytokine receptor, a type I IL receptor, a type II IL receptor, a chemokine receptor or a tumor necrosis receptor superfamily receptor.

37. A conditionally active binding protein according to embodiments 24-36, wherein the cleavable linker comprises a cleavage site.

38. The binding protein of aspect 37, wherein the cleavage site is recognized by a protease.

39.The method of embodiment 38, wherein the protease cleavage site is conditionally recognized by serine protease, cysteine protease, aspartate protease, threonine protease, glutamic acid protease, metalloproteinase, gelatinase, or asparagine peptide lyase. Active binding protein.

40. The method of aspect 38, wherein the protease cleavage site is cathepsin B, cathepsin C, cathepsin D, cathepsin E, cathepsin K, cathepsin L, kallikrein, hK1, hK10, hK15, plasmin, collagena Type IV collagenase, stromelysin, factor Xa, chymotrypsin-like protease, trypsin-like protease, elastase-like protease, subtilisin-like protease, actinidin, bromelain, calpine , Caspase, caspase-3, Mir1-CP, papain, HIV-1 protease, HSV protease, CMV protease, chymosin, renin, pepsin, matriptase, legumine, plasmexin, nepenthesin, metalloexo Peptidase, metalloendopeptidase, matrix metalloprotease (MMP), MMP1, MMP2, MMP3, MMP8, MMP9, MMP10, MMP11, MMP12, MMP13, MMP14, ADAM10, ADAM17, ADAM12, urokinase plasminogen activator (uPA) , Enterokinase, prostate-specific target (PSA, hK3), interleukin-1β converting enzyme, thrombin, FAP (FAP-α), dipeptidyl peptidase, or dipeptidyl peptidase IV (DPPIV/CD26), II Type transmembrane serine protease (TTSP), neutrophil elastase, cathepsin G, proteinase 3, neutrophil serine protease 4, mast cell chimase, mast cell tryptase, dipeptidyl peptidase and dipeptidyl peptidase IV (DPPIV/ CD26), a conditionally active binding protein.

41.The method of embodiment 24, further comprising a half-life extension domain bound to the binding moiety, wherein the half-life extension domain provides a safety switch to the binding protein, and upon cleavage of the linker the binding protein is from the cytokine to the binding moiety and A binding protein that is conditionally active, which is activated by separation of a half-life extension domain, whereby the binding protein is separated from a safety switch.

42. The binding protein of aspect 41, wherein cleavage of the linker is in a tumor microenvironment.

43.A conditionally active binding protein comprising a binding moiety that binds to a cytokine via a non-CDR loop within the binding moiety, wherein the binding moiety is further linked to a half-life extension domain and comprises a cleavable linker, , Wherein the binding protein has an extended half-life prior to its activation by cleavage of the linker, and upon activation, the binding moiety and the half-life extension domain are separated from the cytokine, where in the activated state, the binding protein has an extended half-life. No, conditionally active binding protein.

44. The binding protein of embodiment 43, wherein cleavage of the linker is in the tumor microenvironment.

In vivo half-life prolonging factor

Preferably, the chimeric polypeptide comprises an in vivo half-life extending element. Increasing the in vivo half-life of a therapeutic molecule with a naturally short half-life allows for a more acceptable and manageable dosage regimen without sacrificing the effect. As used herein, a “half-life extending element” increases half-life in vivo, e.g., its size, shape, hydrodynamic radius, charge, or absorption (eg, to be above the renal filtration cutoff), It is part of a chimeric polypeptide that improves pK by altering the parameters of biodistribution, metabolism and elimination. An exemplary method of improving the pK of a polypeptide is by expression of urea in the polypeptide chain that binds to receptors recycled to the plasma membrane of the cell rather than being degraded in lysosomes such as the FcRn receptor and transferrin receptor on endothelial cells. The three types of proteins, such as human IgG, HSA (or fragment), and transferrin, last much longer in human serum than predicted by just their size alone, which is their ability to bind to receptors that are recycled rather than degraded in lysosomes. Is a function of. These proteins or fragments thereof that retain FcRn binding are routinely linked to other polypeptides to extend their serum half-life. In one embodiment, the half-life extending element is a human serum albumin (HSA) binding domain. HSA (SEQ ID NO: 2) can also be linked directly to the pharmaceutical composition or via a short linker. Fragments of HSA can also be used. HSA and fragments thereof can function as both blocking moieties and half-life extending elements. Human IgG and Fc fragments can also perform similar functions.

The serum half-life extending element may also be an antigen-binding polypeptide that binds to a protein having a long serum half-life such as serum albumin, transferrin, and the like. Examples of such polypeptides are polyclonal antibodies, recombinant antibodies, human antibodies, humanized antibodies, single chain variable fragments (scFv), single-domain antibodies such as heavy chain variable domain (VH), light chain variable domain (VL) and camel-type It includes antibodies and fragments thereof, including the variable domain (VHH) of a nanobody, dAb, and the like. Other suitable antigen-binding domains are non-immunoglobulin proteins that mimic antibody binding and/or structure, such as anticalin, apilin, apibody molecule, apimer, afitin, alphabody, abimer, DARPin, phinomer. , Kunitz domain peptides, monobodies and binding domains based on other engineered scaffolds such as SpA, GroEL, fibronectin, lipocalin and CTLA4 scaffolds. Further examples of antigen-binding polypeptides include ligands for the desired receptor, ligand-binding portions of the receptor, lectins, and peptides that bind or link to one or more target antigens.

Some preferred serum-half-life extending elements are polypeptides comprising a complementarity determining region (CDR) and optionally a non-CDR loop. Advantageously, such serum half-life extending elements can extend the serum half-life of cytokines, and also as inhibitors of cytokines (e.g., through steric blocking, non-covalent interactions or combinations thereof) and/or targeting. Can function as a domain. In some examples, the serum half-life extending element is a domain derived from an immunoglobulin molecule (Ig molecule) or engineered protein scaffold that mimics antibody structure and/or binding activity. Ig can be of any class or subclass (IgG1, IgG2, IgG3, IgG4, IgA, IgE, IgM, etc.). The polypeptide chain of the Ig molecule is folded into a series of parallel beta strands linked by loops. In the variable region, three of the loops constitute a "complementarity determining region" (CDR), which determines the antigen binding specificity of a molecule. The IgG molecule comprises at least two heavy chains (H) and two light chains (L) or antigen binding fragments thereof that are interconnected by disulfide bonds. Each heavy chain is composed of a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region. The heavy chain constant region consists of three domains, CH1, CH2 and CH3. Each light chain is composed of a light chain variable region (abbreviated herein as VL) and a light chain constant region. The light chain constant region consists of one domain CL. The VH and VL regions are complementary in sequence, interspersed with more conserved regions, referred to as framework regions (FR), which are hypervariable in sequence and/or are involved in antigen recognition and/or form a commonly structurally defined loop. It can be further subdivided into regions of hypervariability called crystalline regions (CDRs). Each VH and VL consists of 3 CDRs and 4 FRs arranged in the following order from amino-terminus to carboxy-terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. In some embodiments of the present disclosure, at least some or all of the amino acid sequences of FR1, FR2, FR3 and FR4 are part of a “non-CDR loop” of a binding moiety described herein. As shown in FIG. 5 , there are several beta strands arranged in two sheets in the variable domain of the immunoglobulin molecule. The variable domains of the light and heavy chain immunoglobulin chains contain three hypervariable loops or complementarity determining regions (CDRs). The three CDRs of the V domain (CDR1, CDR2, CDR3) are clustered at one end of the beta barrel. CDRs are the loops connecting the beta strands BC, C'-C" and FG of the immunoglobulin fold, while the lower loop connecting the beta strands AB, CC', C"-D and EF of the immunoglobulin fold and the immunoglobulin fold The upper loop connecting the DE strands is a non-CDR loop. In some embodiments of the present disclosure, at least some amino acid residues of the constant domain, which are CH1, CH2, or CH3, are part of a “non-CDR loop” of a binding moiety described herein. Non-CDR loops, in some embodiments, comprise the AB, CD, EF and DE loops of the C1-set domain of an Ig or Ig-like molecule; AB, CC', EF, FG, BC and EC' loops of the C2-set domain of an Ig or Ig-like molecule; One or more of the DE, BD, GF, A(A1A2)B and EF loops of the I (middle)-set domain of an Ig or Ig-like molecule.

Within the variable domain, CDRs are believed to be responsible for antigen recognition and binding, while FR residues are considered scaffolds for CDRs. However, in certain cases some FR residues play an important role in antigen recognition and binding. Framework region residues that affect Ag binding fall into two categories. The first are the FR residues that contact the antigen, and thus are part of the binding-site and some of these residues are close to the CDRs in sequence. Other residues are those that are far from the CDR in sequence, but are close in the three-dimensional structure of the molecule, for example loops in the heavy chain. The serum half-life extension domain (eg, a domain comprising a CDR) may comprise at least one non-CDR loop. In some embodiments, the non-CDR loop provides a binding site for binding cytokines, bulk serum proteins, or other target antigens.

In addition to or alternatively to containing the CDRs, the serum half-life extending element comprises a non-CDR loop. In some embodiments, the non-CDR loop is modified to create an antigen binding site specific for a desired target antigen, such as a bulk serum protein such as albumin, or a cytokine moiety or other target antigen. It is contemplated that various techniques can be used to modify non-CDR loops, for example site-directed mutagenesis, random mutagenesis, insertion of at least one amino acid foreign to the non-CDR loop amino acid sequence, amino acid substitutions. do. The antigenic peptide is inserted in the non-CDR loop in some instances. In some instances, antigenic peptides replace non-CDR loops. The modification to create an antigen binding site is sometimes only in one non-CDR loop. In other cases, more than one non-CDR loop is modified. For example, the variant is in any one of the non-CDR loops shown in Figure 5 , namely AB, CC', C"D, EF and DE. In some cases the variant is in the DE loop. In other cases the variant is in the DE loop. Is in all four of the AB, CC', C"-D, EF loops.

In some instances, the serum half-life extending element contains a CDR that has double binding specificity and specifically binds to bulk serum proteins such as serum albumin and a non-CDR loop that specifically binds and blocks cytokine domains. In another example, the serum half-life extending element contains a CDR that specifically binds to a target antigen, such as a cytokine domain or other target antigen, and a non-CDR loop that specifically binds to a bulk serum protein, such as serum albumin. Preferably, the serum half-life extending element inhibits the binding of the cytokine domain to the cognate cytokine receptor, for example through steric occlusion, through specific intermolecular interactions, or a combination of the two.

In some embodiments, the serum half-life extending element non-covalently binds directly to a cytokine and inhibits its activity.

In certain instances, the binding moiety binds to a cytokine through one or more of the AB, CC', C" D and EF loops and to a bulk serum protein such as albumin through one or more of the BC, C'C" and FG loops. Combine. In certain instances, the binding moiety binds to a bulk serum protein such as albumin through an AB, CC', C" D or EF loop and to a cytokine through its BC, C'C" or FG loop. In certain instances, the binding moiety binds to a bulk serum protein such as albumin through its AB, CC', C" D and EF loops and to cytokines through its BC, C'C" and FG loops. In certain instances, the binding moiety binds to a bulk serum protein such as albumin through one or more of the AB, CC', C" D and EF loops and to cytokines through one or more of the BC, C'C" and FG loops. Combine.

The binding moiety is any kind of polypeptide. For example, in certain cases the binding moiety is a natural peptide, a synthetic peptide or a fibronectin scaffold or an engineered bulk serum protein. Bulk serum proteins include, for example, albumin, fibrinogen or globulin. In some embodiments, the binding moiety is an engineered scaffold. Engineered scaffolds are, for example, sdAb, scFv, Fab, VHH, fibronectin type III domains, immunoglobulin-like scaffolds (Halaby et al., 1999. Prot Eng 12(7):563-571 as suggested in), DARPin, cystine knotted peptide, lipocalin, three-helix bundle scaffold, protein G-related albumin-binding module, or DNA or RNA aptamer scaffold.

In some cases, the serum half-life extending element binds to the cytokine domain through its non-CDR loop and the cytokine domain is further linked to the targeting domain as described herein. In some cases, the serum half-life extending element comprises a binding site for a bulk serum protein. In some embodiments, the CDRs provide a binding site for bulk serum proteins. The bulk serum protein is in some instances globulin, albumin, transferrin, IgG1, IgG2, IgG4, IgG3, IgA monomer, factor XIII, fibrinogen, IgE or pentameric IgM. In some embodiments, the CDRs form a binding site for an immunoglobulin light chain, such as an IgK free light chain or an Igλ free light chain.

One exemplary conditionally active protein is shown in FIG. 6 . In the illustrated example, the non-CDR loop in the serum albumin binding domain (eg, dAb) can form a binding site for the cytokine IL-2. In this example, the binding site for serum albumin can be formed by the CDRs of the serum albumin binding domain.

The serum half-life extending element can be any type of binding domain including, but not limited to, domains from monoclonal antibodies, polyclonal antibodies, recombinant antibodies, human antibodies, humanized antibodies. In some embodiments, the binding moiety is a single chain variable fragment (scFv), a single-domain antibody such as a heavy chain variable domain (VH), a light chain variable domain (VL), and a variable domain of a camel-derived nanobody (VHH). In other embodiments, the binding moiety is a non-Ig binding domain, i.e., an antibody mimic, such as anticaline, apilin, apibody molecule, apimer, afitin, alphabody, abimer, DARPin, pinomer, Kunitz Domain peptides and monobodies.

In other embodiments, the serum half-life extending element may be a water soluble polymer or a peptide conjugated to a water soluble polymer such as PEG. “PEG,” “polyethylene glycol” and “poly(ethylene glycol)” as used herein are interchangeable and encompasses any non-peptidyl water soluble poly(ethylene oxide). The term “PEG” also means a polymer containing the majority of the _{—OCH 2} CH _{2 —repeating subunits, ie more than 50%.} With respect to a particular form, PEG may have any of a variety of molecular weights, as well as structures or geometries such as "branched", "linear", "branched", "multifunctional", and the like described in detail below. PEG is not limited to a specific structure and is linear (e.g., end-capped, e.g., alkoxy PEG or difunctional PEG), branched or multi-arm (e.g., branched PEG or polyol attached to the core). PEG), dendritic (or star) architecture, each with or without one or more degradable linkages. Moreover, the internal structure of PEG can be organized into any number of different repeating patterns and is selected from the group consisting of homopolymers, alternating copolymers, random copolymers, block copolymers, alternating terpolymers, random terpolymers and block terpolymers. Can be. PEG can be conjugated to polypeptides and peptides through any suitable method. Typically reactive PEG derivatives such as N-hydroxysuccinamidyl ester PEG are amine, sulfhydryl, carboxylic acid or hydroxyl functional groups such as cysteine, lysine, asparagine, glutamine, theonine, tyrosine, serine, aspartic acid and glutamic acid. It reacts with a peptide or polypeptide containing an amino acid having a side chain containing

Targeting and Conservation Domain

For certain applications, it may be desirable to maximize the amount of time the construct is in a desired location on the body. This can be achieved by including one additional domain in the chimeric polypeptide (fusion protein) to influence its movement within the body. For example, a chimeric nucleic acid can encode a domain that directs the polypeptide to a location in the body, such as a tumor cell or a site of inflammation; This domain is referred to as a “targeting domain” and/or encodes a domain that carries the polypeptide at a location in the body, eg, a tumor cell or site of inflammation; This domain is referred to as a “conservation domain”. In some embodiments, a domain can function as both a targeting and a retention domain. In some embodiments, the targeting domain and/or the retention domain is specific for a protease-rich environment. In some embodiments, the encoded targeting domain and/or retention domain is specific for regulatory T cells (Tregs), e.g. targeting CCR4 or CD39 receptors. Other suitable targeting and/or retention domains include those with cognate ligands that are overexpressed in inflamed tissue, such as the IL-1 receptor or the IL-6 receptor. In other embodiments, suitable targeting and/or retention domains include those with cognate ligands that are overexpressed in tumor tissue, such as Epcam, CEA or mesothelin. In some embodiments, the targeting domain is linked to the interleukin through a linker that is cleaved at the site of action (eg, by an inflammation or cancer specific protease) to release the interleukin full activity at the desired site. In some embodiments, the targeting and/or retention domain is linked to the interleukin through a linker that is not cleaved at the site of action (eg, by inflammation or cancer specific proteases), allowing the cytokine to remain at the desired site.

The selected antigen is in some cases expressed on the surface of diseased cells or tissues, such as tumors or cancer cells. Antigens useful for tumor targeting and maintenance include, but are not limited to, EpCAM, EGFR, HER-2, HER-3, c-Met, FOLR1 and CEA. The pharmaceutical compositions disclosed herein also include proteins comprising two targeting and/or retention domains that bind to two different target antigens known to be expressed in diseased cells or tissues. Exemplary pairs of antigen binding domains include, but are not limited to, EGFR/CEA, EpCAM/CEA and HER-2/HER-3.

Suitable targeting and/or retention domains include polyclonal antibodies, recombinant antibodies, human antibodies, humanized antibodies, single chain variable fragments (scFv), single-domain antibodies such as heavy chain variable domain (VH), light chain variable domain (VL) and camel -Includes antigen-binding domains such as antibodies and fragments thereof, including variable domains (VHH) of type nanobody, dAb, etc. Other suitable antigen-binding domains are non-immunoglobulin proteins that mimic antibody binding and/or structure, such as anticalin, apilin, apibody molecule, apimer, afitin, alphabody, abimer, DARPin, pinomer, Binding domains based on Kunitz domain peptides, monobodies and other engineered scaffolds such as SpA, GroEL, fibronectin, lipocalin and CTLA4 scaffolds. Additional examples of antigen-binding polypeptides include ligands for the desired receptor, ligand-binding portions of the receptor, lectins, and peptides that bind or associate with one or more target antigens.

In some embodiments, the targeting and/or retention domain specifically binds to a cell surface molecule. In some embodiments, the targeting and/or retention domain specifically binds to a tumor antigen. In some embodiments, the targeting polypeptide is fibroblast activating protein alpha (FAPa), trophoblast glycoprotein (5T4), tumor-associated calcium signal transducer 2 (Trop2), fibronectin EDB (EDB-FN), fibronectin EIIIB domain, It specifically and independently binds to a tumor antigen selected from at least one of CGS-2, EpCAM, EGFR, HER-2, HER-3, cMet, CEA and FOLR1. In some embodiments, the targeting polypeptide specifically and independently binds two different antigens, wherein at least one of the antigens is fibroblast activating protein alpha (FAPa), trophoblast glycoprotein (5T4), tumor-associated calcium. Signal transducer 2 (Trop2), fibronectin EDB (EDB-FN), fibronectin EIIIB domain, CGS-2, EpCAM, EGFR, HER-2, HER-3, cMet, CEA and FOLR1.

The targeting and/or retaining antigen may be a tumor antigen expressed on a tumor cell. Tumor antigens are well known in the art and, for example, EpCAM, EGFR, HER-2, HER-3, c-Met, FOLR1, PSMA, CD38, BCMA and CEA. 5T4, AFP, B7-H3, Cadherin-6, CAIX, CD117, CD123, CD138, CD166, CD19, CD20, CD205, CD22, CD30, CD33, CD352, CD37, CD44, CD52, CD56, CD70, CD71, CD74 , CD79b, DLL3, EphA2, FAP, FGFR2, FGFR3, GPC3, gpA33, FLT-3, gpNMB, HPV-16 E6, HPV-16 E7, ITGA2, ITGA3, SLC39A6, MAGE, mesothelin, Muc1, Muc16, NaPi2b, Nectin-4, P-cadherin, NY-ESO-1, PRLR, PSCA, PTK7, ROR1, SLC44A4, SLTRK5, SLTRK6, STEAP1, TIM1, Trop2, WT1.

The targeting and/or retaining antigen can be an immune checkpoint protein. Examples of immune checkpoint proteins are CD27, CD137, 2B4, TIGIT, CD155, ICOS, HVEM, CD40L, LIGHT, TIM-1, OX40, DNAM-1, PD-L1, PD1, PD-L2, CTLA-4, CD8. , CD40, CEACAM1, CD48, CD70, A2AR, CD39, CD73, B7-H3, B7-H4, BTLA, IDO1, IDO2, TDO, KIR, LAG-3, TIM-3, or VISTA. .

The targeting and/or retaining antigen can be a cell surface molecule such as a protein, lipid or polysaccharide. In some embodiments, the targeting and/or retaining antigen is a tumor cell, a virus infected cell, a bacterial infected cell, an damaged red blood cell, an arterial plaque cell, an inflammatory or fibrous tissue cell. Targeting and/or retaining antigens can include immune response modulators. Examples of immune response modulators include granulocyte-macrophage colony stimulating factor (GM-CSF), macrophage colony stimulating factor (M-CSF), granulocyte colony stimulating factor (G-CSF), interleukin 2 (IL-2), interleukin 3 (IL-3), interleukin 12 (IL-12), interleukin 15 (IL-15), B7-1 (CD80), B7-2 (CD86), GITRL, CD3 or GITR.

The targeting and/or retaining antigen can be a cytokine receptor. Examples of cytokine receptors include type I cytokine receptors such as GM-CSF receptor, G-CSF receptor, type I IL receptor, Epo receptor, LIF receptor, CNTF receptor, TPO receptor; Type II cytokine receptors such as IFN-alpha receptors (IFNAR1, IFNAR2), IFB-beta receptors, IFN-gamma receptors (IFNGR1, IFNGR2), type II IL receptors; Chemokine receptors such as CC chemokine receptor, CXC chemokine receptor, CX3C chemokine receptor, and XC chemokine receptor; Tumor necrosis receptor superfamily receptors such as TNFRSF5/CD40, TNFRSF8/CD30, TNFRSF7/CD27, TNFRSF1A/TNFR1/CD120a, TNFRSF1B/TNFR2/CD120b; TGF-beta receptors such as TGF-beta receptor 1 and TGF-beta receptor 2; IL-1 receptor, CSF-1R, PDGFR (PDGFRA, PDGFRB), including, but not limited to, Ig superfamily receptors such as SCFR.

Linker

As mentioned above, the pharmaceutical composition comprises one or more linker sequences. The linker sequence serves to provide flexibility between the polypeptides so that, for example, the blocking moiety can inhibit the activity of the cytokine polypeptide. The linker sequence may be located between any or all of the cytokine polypeptide, serum half-life extending element and/or blocking moiety. As described herein, at least one of the linkers is protease cleavable and contains (one or more) cleavage sites for the (one or more) desired proteases. Preferably, the desired protease is enriched or selectively expressed at the desired cytokine active site (eg, tumor microenvironment). Thus, the fusion protein is preferentially or selectively cleaved at the desired cytokine active site.

Suitable linkers are 3 amino acids, including 1 amino acid (e.g. Gly) to 20 amino acids, 2 amino acids to 15 amino acids, 4 amino acids to 10 amino acids, amino acids to 9 amino acids, 6 amino acids to 8 amino acids, or 7 amino acids to 8 amino acids. Can be of different lengths, such as to 12 amino acids, and 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60 amino acids.

It is recognized that the orientation of the components of the pharmaceutical composition is primarily a matter of design choice and multiple orientations are possible and all are intended to be encompassed by the present disclosure. For example, the blocking moiety can be located C-terminally or N-terminally to the cytokine polypeptide.

Proteases known to be associated with diseased cells or tissues include serine protease, cysteine protease, aspartate protease, threonine protease, glutamic acid protease, metalloprotease, asparagine peptide lyase, serum protease, cathepsin, cathepsin B, cathepsin C, ca. Thepsin D, cathepsin E, cathepsin K, cathepsin L, kallikrein, hK1, hK10, hK15, plasmin, collagenase, type IV collagenase, stromelysin, factor Xa, chymotrypsin-like protease, Trypsin-like protease, elastase-like protease, subtilisin-like protease, actinidain, bromelain, calpain, caspase, caspase-3, Mirl-CP, papain, HIV-1 protease, HSV Protease, CMV protease, chymosin, renin, pepsin, matriptase, legumine, plasmexin, nepenthesin, metalloexopeptidase, metalloendopeptidase, matrix metalloprotease (MMP), MMP1, MMP2, MMP3, MMP8, MMP9, MMP13, MMP11, MMP14, urokinase plasminogen activator (uPA), enterokinase, prostate-specific antigen (PSA, hK3), interleukin-1β converting enzyme, thrombin, FAP (FAP-a) , Dipeptidyl peptidase, mephrin, granzyme and dipeptidyl peptidase IV (DPPIV/CD26). Proteases capable of cleaving the amino acid sequence encoded by the chimeric nucleic acid sequence provided herein are, for example, prostate specific antigen (PSA), matrix metalloproteinase (MMP), A disintigrin and metalloproteina. Agents (ADAM), plasminogen activators, cathepsins, caspases, tumor cell surface proteases and elastases. The MMP can be, for example, matrix metalloproteinase 2 (MMP2) or matrix metalloproteinase 9 (MMP9).

Proteases useful in the methods disclosed herein are shown in Table 1, and exemplary proteases and their cleavage sites are shown in Table 1A:

Pharmaceutical compositions comprising a polypeptide sequence are provided herein. It is understood that, as with all peptides, polypeptides and proteins, including fragments thereof, further modifications (amino acid sequence variants) may occur in the amino acid sequence of the peptide, polypeptide or chimeric polypeptide that does not alter the properties or function of the protein. . These modifications include conservative amino acid substitutions and are discussed in more detail below.

The compositions provided herein have a desired function. The composition consists of at least an IL-2 polypeptide, a blocking moiety, such as a steric blocking polypeptide, and an optional serum half-life extending element, and an optional targeting polypeptide, and at least one linker connecting each polypeptide in the composition Has. The first polypeptide, for example the IL-2 mutein, is provided to be an active agent. Blocking moieties are provided to block the activity of the interleukin. Linker polypeptides, such as protease cleavable polypeptides, are provided to be cleaved by a protease that is specifically expressed at the intended target of the active agent. Optionally, the blocking moiety blocks the activity of the first polypeptide by binding to the interleukin polypeptide. In some embodiments, a blocking moiety, for example. The steric blocking peptide is linked to the interleukin via a protease-cleavable linker that is cleaved at the site of action (eg, by an inflammation-specific or tumor-specific protease) to release the cytokine total activity at the desired site.

The protease cleavage site may be a naturally occurring protease cleavage site or an artificially engineered protease cleavage site. The artificially engineered protease cleavage site can be cleaved by more than one protease specific for the desired environment in which cleavage will occur, for example a tumor. The protease cleavage site can be cleaved by at least one protease, at least two proteases, at least three proteases, or at least four proteases.

In some embodiments, the linker is a glycine-glycine, a sortase-recognition motif, or a sortase-recognition motif and a peptide sequence (Gly ₄ Ser) _n (SEQ ID NO: 126) or (Gly ₃ Ser) _n (SEQ ID NO: 127. ), where n is 1, 2, 3, 4 or 5. In one embodiment, the sortase-recognition motif comprises the peptide sequence LPXTG (SEQ ID NO: 125), wherein X is any amino acid. In one embodiment, the covalent bond is between a reactive lysine residue attached to the C-terminus of the cytokine polypeptide and a reactive aspartic acid attached to the N-terminus of a blocking or other moiety. In one embodiment, the covalent bond is between a reactive aspartic acid moiety attached to the N-terminus of the cytokine polypeptide and a reactive lysine moiety attached to the C-terminus of a blocking or other moiety.

Cleavage and induction

As described herein, the activity of cytokine polypeptides in the context of fusion proteins is weakened, and protease cleavage at the desired active site, such as in the tumor microenvironment, is from fusion proteins that are much more active as cytokine receptor agonists than fusion proteins. Releases a form of cytokine. For example, the cytokine-receptor activation (agonist) activity of the fusion polypeptide is at least about 10X, at least about 50X, at least about 100X, at least about 250X, than the cytokine receptor activation activity of the cytokine polypeptide as a separate molecular entity, May be at least about 500X, or at least about 1000X smaller. A cytokine polypeptide that is part of a fusion protein is a separate molecular entity when it contains an amino acid that is substantially identical to the cytokine polypeptide and does not contain substantially any additional amino acids and is not bound (by covalent or non-covalent bonds) with other molecules. exist. If desired, as a separate molecular entity, the cytokine polypeptide may contain some additional amino acid sequence, such as a tag or short sequence, to aid in expression and/or purification.

In another example, the cytokine-receptor activation (agonist) activity of the fusion polypeptide is at least about 10X greater than the cytokine receptor activation activity of the polypeptide containing the cytokine polypeptide produced by cleavage of a protease cleavable linker in the fusion protein. , At least about 50X, at least about 100X, at least about 250X, at least about 500X, or about 1000X less. In other words, the cytokine receptor activation (agonist) activity of the polypeptide containing the cytokine polypeptide produced by cleavage of a protease cleavable linker in the fusion protein is at least about 10X, at least about 10 times that of the fusion protein. 50X, at least about 100X, at least about 250X, at least about 500X, or at least about 1000X greater.

Polypeptide substitution

The polypeptides described herein may have the same amino acid sequence of the corresponding naturally occurring protein (e.g., IL-2, IL-15, HSA) or have an amino acid sequence different from the naturally occurring protein as long as the desired function is maintained. Components (eg, cytokines, blocking moieties). It is understood that one way to define any known modifications and derivatives of the disclosed proteins and nucleic acids encoding them, or what may occur, is through defining sequence variants in terms of identity to a particular known reference sequence. In particular at least 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89 for chimeric polypeptides provided herein. , 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 percent identity of a polypeptide or nucleic acid is disclosed. For example, at least 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85 for the sequence of any nucleic acid or polypeptide provided herein. , 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 percent identity of a polypeptide or nucleic acid is provided. One of skill in the art readily understands how to determine the identity of two polypeptides or two nucleic acids. For example, identity can be calculated after aligning the two sequences so that their highest level of identity is achieved.

Another way to calculate identity can be done by a published algorithm. The optimal sequence alignment for comparison is Smith and Waterman Adv. Appl. Math. 2:482 (1981) by the local identity algorithm, Needleman and Wunsch, J. Mol. Biol. 48:443 (1970) by the identity alignment algorithm, Pearson and Lipman, Proc. Natl. Acad. Sci. USA 85: 2444 (1988) computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, Wis.) or by inspection.

See, for example, Zuker, Science 244:48-52 (1989); Jaeger et al., Proc. Natl. Acad. Sci. USA 86:7706-7710 (1989); Jaeger et al., Methods Enzymol. The same type of identity can be obtained for nucleic acids by the algorithm disclosed in 183:281-306 (1989), which is incorporated herein by reference, at least for materials related to nucleic acid alignment. While it is understood that any method may typically be used and the results of these various methods may differ in certain instances, the skilled artisan is said to have found identity with at least one of these methods, and that the sequence is said to have the specified identity. Understand whether it can be disclosed in.

Protein modification includes amino acid sequence modification. Modifications in the amino acid sequence can occur naturally as allelic variations (e.g., due to genetic polymorphism), can occur due to environmental influences (e.g., due to exposure to UV light), or human Intervention (eg, by mutagenesis of the cloned DNA sequence), such as induced point, deletion, insertion and substitution mutations. These modifications can result in changes in the amino acid sequence, provide silent mutations, modify restriction sites, or provide other specific mutations. Amino acid sequence modifications typically belong to one or more of the three classes of substitution, insertion or deletion modifications. Insertions include amino and/or carboxyl terminal fusions as well as intrasequence insertions of single or multiple amino acid residues. Insertions will generally be smaller than those of amino or carboxyl terminal fusions, for example on the order of 1 to 4 residues. Deletion is characterized by the removal of one or more amino acid residues from the protein sequence. Typically, no more than about 2 to 6 residues are deleted at any one site within the protein molecule. Amino acid substitutions are typically of a single residue, but can occur at many different positions at once; Insertions will generally be on the order of about 1 to 10 amino acid residues; The deletion will range from about 1 to 30 residues. Deletion or insertion preferably consists of contiguous pairs, i.e. deletion of 2 residues or insertion of 2 residues. Substitutions, deletions, insertions or any combination thereof can be combined to arrive at the final construct. The mutation should not place the sequence outside the reading frame and preferably will not create a complementary region capable of producing a secondary mRNA structure. Substitutional modification is that at least one residue is removed and another residue is inserted at that position. Such substitutions are generally made according to Table 2 below and are referred to as conservative substitutions.

Modifications, including specific amino acid substitutions, are made by known methods. For example, the modification is made by site-specific mutagenesis of the nucleotides in the DNA encoding the polypeptide, thereby generating the DNA encoding the modification, and then expressing the DNA in recombinant cell culture. Techniques for making substitutional mutations at predetermined sites in DNA having a known sequence, such as M13 primer mutagenesis and PCR mutagenesis, are well known.

Variations can be chosen to optimize the binding. For example, affinity maturation techniques can be used to alter the binding of scFvs by introducing random mutations inside the complementarity determining region (CDR). These random mutations can be introduced using a variety of techniques including radiation, chemical mutation or mistake-prone PCR. For example, multiple rounds of mutations and selection can be performed using phage display.

The present disclosure also relates to nucleic acids encoding the chimeric polypeptides described herein, and to the use of such nucleic acids to generate chimeric polypeptides and for therapeutic purposes. For example, the present invention includes DNA and RNA molecules encoding chimeric polypeptides (eg, mRNA, self-replicating RNA) and the therapeutic use of such DNA and RNA molecules.

Exemplary composition

Exemplary fusion proteins of the invention combine the elements described above in various orientations. The directions described in this section are meant by way of example and are not to be considered limiting.

In some embodiments, the fusion protein comprises an IL-2 polypeptide, a blocking moiety and a half-life extending element. In some embodiments, the IL-2 polypeptide is located between a half-life extending element and a blocking moiety. In some embodiments, the IL-2 polypeptide is N-terminal to the blocking moiety and half-life extending element. In some such embodiments, the IL-2 polypeptide is proximal to a blocking moiety; In some such embodiments, the IL-2 polypeptide is proximal to a half-life extending element. At least one protease-cleavable linker should be included in all embodiments so that the IL-2 polypeptide is active upon cleavage. In some embodiments, the IL-2 polypeptide is C-terminal to the blocking moiety and half-life extending element. The additional elements may be attached to each other by cleavable linkers, non-cleavable linkers or direct fusion. In some cases it is beneficial to include two identical cytokines to promote dimerization.

In some embodiments, the blocking domain used can extend half-life, and the IL-2 polypeptide is located between two such blocking domains. In some embodiments, the IL-2 polypeptide is located between two blocking domains, one of which is capable of extending half-life.

In some embodiments, two cytokines are included in the same construct, and at least one is IL-2. In some embodiments, each cytokine is linked to two blocking domains (a total of three in one molecule), having a blocking domain between the two cytokine domains. In some embodiments, one or more additional half-life extension domains may be included to optimize pharmacokinetic properties.

In some embodiments, three cytokines are included in the same construct. In some embodiments, the third cytokine may function to block the other two instead of the blocking domain between the two cytokines.

Preferred half-life extending elements for use in the fusion protein are human serum albumin (HSA), an antibody or antibody fragment that binds serum albumin (e.g., scFV, dAb), human or humanized IgG, or any of the foregoing. Is a short story. In some preferred embodiments, the blocking moiety is human serum albumin (HSA), or an antibody or antibody fragment that binds serum albumin, binds to a cytokine and prevents activation or activation of a cytokine receptor, another cytokine. It is an antibody or a fragment of any of the foregoing. In a preferred embodiment comprising an additional targeting domain, the targeting domain is an antibody that binds to cell surface proteins enriched on the surface of cancer cells such as EpCAM, FOLR1 and fibronectin.

Treatment method and pharmaceutical composition

In addition, the risk of having or developing a disease or disorder, such as a proliferative disease, neoplastic disease, inflammatory disease, immune disease, autoimmune disease, infectious disease, viral disease, allergic reaction, parasitic reaction, or graft-versus-host disease Methods of treating a subject with a disorder are provided. A method of administering to a subject in need thereof an effective amount of a fusion protein as disclosed herein, typically administered as a pharmaceutical composition. In some embodiments, the method further comprises selecting a subject who has or is at risk of developing such disease or disorder. The pharmaceutical composition preferably comprises a blocked cytokine, fragment or mutein thereof that is activated at the site of inflammation or tumor. In one embodiment, the chimeric polypeptide comprises a cytokine polypeptide, a fragment or mutein thereof and a serum half-life extending element. In another embodiment, the chimeric polypeptide comprises a cytokine polypeptide, a fragment or mutein thereof and a blocking moiety, e.g., a steric blocking polypeptide, wherein the steric blocking polypeptide is a cytokine polypeptide, a fragment thereof. Alternatively, the mutein activity can be sterically blocked. In another embodiment, the chimeric polypeptide comprises a cytokine polypeptide, a fragment or mutein thereof, a blocking moiety and a serum half-life extending element.

Inflammation is part of a complex biological response of body tissues to harmful stimuli such as pathogens, damaged cells or irritants, and is a protective response associated with immune cells, blood vessels and molecular mediators. The function of inflammation is to eliminate the initial cause of cell damage, to clear out damaged necrotic cells and tissues from the original injury and inflammatory process, and to initiate tissue repair. Inflammation can be caused by symptoms or diseases, such as infections such as cancer, atherosclerosis, allergies, myopathy, HIV, obesity or an autoimmune disease. Autoimmune disease is a chronic condition that results from an abnormal immune response to an auto-antigen. Autoimmune diseases that can be treated with the polypeptides disclosed herein include, but are limited to, lupus, celiac disease, diabetes mellitus type 1, Graves disease, inflammatory bowel disease, multiple sclerosis, psoriasis, rheumatoid arthritis, and systemic lupus erythematosus. It doesn't work.

The pharmaceutical composition may comprise one or more protease-cleavable linker sequences. The linker sequence serves to provide flexibility between the polypeptides so that each polypeptide can inhibit the activity of the first polypeptide. The linker sequence may be located between any or all of the cytokine polypeptide, fragment or mutein thereof, a blocking moiety and a serum half-life extending element. Optionally, the composition comprises 2, 3, 4 or 5 linker sequences. The linker sequence, which is 2, 3 or 4 linker sequences, can be the same or different linker sequences. In one embodiment, the linker sequence comprises GGGGS (SEQ ID NO: 132), GSGSGS (SEQ ID NO: 133) or G(SGGG) ₂ SGGT (SEQ ID NO: 134). In another embodiment, the linker is the group consisting of HSSKLQ (SEQ ID NO: 25), GPLGVRG (SEQ ID NO: 128), IPVSLRSG (SEQ ID NO: 129), VPLSLYSG (SEQ ID NO: 130) and SGESPAYYTA (SEQ ID NO: 131). It includes a protease-cleavable sequence selected from.

In some embodiments, the linker is kallikrein, thrombin, chimase, carboxypeptidase A, cathepsin G, elastase, PR-3, granzyme M, calpine, matrix metalloproteinase (MMP), It is cleaved by a protease selected from the group consisting of a plasminogen activator, cathepsin, caspase, tryptase or tumor cell surface protease.

Suitable linkers are 3 amino acids, including 1 amino acid (e.g. Gly) to 20 amino acids, 2 amino acids to 15 amino acids, 4 amino acids to 10 amino acids, amino acids to 9 amino acids, 6 amino acids to 8 amino acids, or 7 amino acids to 8 amino acids. It can be of different lengths, such as from 12 amino acids to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 , 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45 , 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60 amino acids.

Further provided are methods of treating a subject having or at risk of developing cancer. The method comprises administering an effective amount of a chimeric polypeptide (fusion protein) as disclosed herein, typically administered as a pharmaceutical composition, to a subject in need thereof. In some embodiments, the method further comprises selecting a subject who has or is at risk of developing cancer. The pharmaceutical composition preferably comprises a blocked cytokine, fragment or mutein thereof that is activated at the site of the tumor. Preferably the tumor is a solid tumor. The cancer may be colon cancer, lung cancer, melanoma, sarcoma, renal cell carcinoma, and breast cancer, but is not limited thereto.

The method includes chemotherapeutic agents (e.g., adriamycin, serubidin, bleomycin, alkeran, belban, oncobin, fluororacil, thiotepa, methotrexate, bisantrene, presbyotrope, thiguanine, cytaribine, Procaravigin), immune-tumor agents (e.g. anti-PD-L1, anti-CTLA4, anti-PD-1, anti-CD47, anti-GD2), cell therapy (e.g., CAR-T, T-cell therapy), oncolytic viruses, and the like, may further include administration of one or more additional agents to treat cancer.

Pharmaceutical formulations or compositions containing a chimeric polypeptide and a pharmaceutically acceptable carrier are provided herein. The compositions provided herein are suitable for in vitro or in vivo administration. A pharmaceutically acceptable carrier means a substance that is biologically or otherwise undesirable, i.e., the substance causes undesirable biological effects or interacts in a deleterious manner with other components of the pharmaceutical formulation or composition in which it is contained. It is administered to the subject without having to do so. The carrier is chosen to minimize the degradation of the active ingredient and minimize side effects in the subject.

Suitable carriers and their formulations are described in Remington: The Science and Practice of Pharmacy, 21 ^st Edition, David B. Troy, ed., Lippicott Williams & Wilkins (2005). Typically, the formulation may be of high rigidity or storage if desired, but an appropriate amount of a pharmaceutically-acceptable salt is used in the formulation to render the formulation isotonic. Examples of pharmaceutically-acceptable carriers include, but are not limited to, sterile water, saline, buffered solutions such as Ringer's solution, and dextrose solutions. The pH of the solution is generally about 5 to about 8 or about 7 to 7.5. Other carriers include sustained release formulations such as semipermeable matrices of solid hydrophobic polymers containing immunogenic polypeptides. The matrix can be in the form of a molded article such as a film, liposome or microparticle. For example, certain carriers may be more desirable depending on the route of administration and the concentration of the composition being administered. Carriers are those suitable for administration of a chimeric polypeptide or a nucleic acid sequence encoding a chimeric polypeptide to a human or other subject.

The pharmaceutical formulation or composition is administered in a variety of ways depending on whether local or systemic treatment is desired and the site being treated. The composition may be any of several including topical, oral, parenteral, intravenous, intra-articular, intraperitoneal, intramuscular, subcutaneous, intracavity, transdermal, intrahepatic, intracranial, spray/inhalation, or installation via bronchoscopy. It is administered via the route of administration. In some embodiments, the composition is administered topically (non-systemic), including intratumorally, intra-articular, intrathecal, and the like.

Formulations for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions and emulsions. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil and injectable organic esters such as ethyl oleate. Aqueous carriers include water, alcoholic/aqueous solutions, emulsions or suspensions, including saline and buffered media. Parenteral vehicles include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, Ringer's lactate or fixed oil. Intravenous vehicles include fluid and nutritional supplements, electrolyte supplements (eg, based on Ringer's dextrose), and the like. Preservatives and other additives such as, for example, antimicrobial agents, anti-oxidants, chelating agents and inert gases, etc. are optionally present.

Formulations for topical administration include ointments, lotions, creams, gels, drops, suppositories, sprays, liquids and powders. Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners and the like are optionally required or preferred.

Compositions for oral administration include powders or granules, suspensions or solutions in water or non-aqueous media, capsules, bags or tables. Thickeners, flavors, diluents, emulsifiers, dispersion aids or binders are optionally preferred.

Optionally, the chimeric polypeptide or nucleic acid sequence encoding the chimeric polypeptide is administered by the vector. There are a number of compositions and methods that can be used to deliver nucleic acid molecules and/or polypeptides to cells in vitro or in vivo , for example via expression vectors. These methods and compositions can be broadly divided into two classes: virus-based delivery systems and non-viral-based delivery systems. Such methods are well known in the art and are readily adaptable for use with the compositions and methods described herein. Such compositions and methods are for transfecting or transducing cells in vitro or in vivo, e.g. for generating cell lines expressing and preferably secreting the encoded chimeric polypeptide or therapeutically delivering nucleic acids to a subject. Can be used for Components of the chimeric nucleic acid disclosed herein are typically operably linked in frame to encode the fusion protein.

As used herein, a plasmid or viral vector is an agent that transfers a disclosed nucleic acid into a cell without degradation, and includes a promoter that produces expression of the nucleic acid molecule and/or polypeptide in the cell to which it is delivered. Viral vectors are, for example, adenovirus, adeno-associated virus, herpes virus, vaccinia virus, polio virus, sindbis and other RNA viruses, including those viruses with the HIV backbone. Also preferred are any virus families that share the properties of these viruses making them suitable for use as vectors. In general, retroviral vectors are described in Coffin et al., Retroviruses, Cold Spring Harbor Laboratory Press (1997), which are incorporated herein by reference for vectors and methods of making them. Construction of replication-defective adenoviruses has been described (Berkner et al., J. Virol. 61:1213-20 (1987); Massie et al., Mol. Cell. Biol. 6:2872-83 (1986); Haj-Ahmad et al. , J. Virol. 57:267-74 (1986); Davidson et al., J. Virol. 61:1226-39 (1987); Zhang et al., BioTechniques 15:868-72 (1993)). The advantage and use of these viruses as vectors is that they can replicate within initially infected cells, but cannot form new infectious virus particles, limiting the extent to which they can spread to other cell types. Recombinant adenovirus has been shown to achieve high efficiency after direct in vivo delivery to airway epithelium, hepatocytes, vascular endothelium, CNS parenchyma and many other tissue sites. Other useful systems include, for example, replication and host-restricted non-replicating vaccinia virus vectors.

A provided polypeptide and/or nucleic acid molecule can be delivered via virus like particles. Virus-like particles (VLPs) are composed of viral protein(s) derived from the structural proteins of the virus. Methods of making and using virus-like particles are described, for example, in Garcea and Gissmann, Current Opinion in Biotechnology 15:513-7 (2004).

Provided polypeptides can be delivered by subviral high-density (DB). DB transports proteins into target cells by membrane fusion. Methods of creating and using DBs are described, for example, in Pepperl-Klindworth et al., Gene Therapy 10:278-84 (2003).

Provided polypeptides can be delivered by capsular aggregates. The method of making and using the film agglomerates is described in International Publication No. WO 2006/110728.

Non-viral based delivery methods may include expression vectors comprising nucleic acid molecules and nucleic acid sequences encoding the polypeptide, wherein the nucleic acid is operably linked to an expression control sequence. Suitable vector backbones include those routinely used in the art, such as, for example, plasmids, artificial chromosomes, BAC, YAC or PAC. Numerous vector and expression systems are commercially available from companies such as Novagen (Madison, Wis.), Clonetech (Paal Alto, Calif.), Stratagene (La Jolla, Calif.) and Invitrogen/Life Technologies (Carlsbad, Calif.). Do. Vectors typically contain one or more regulatory regions. Regulatory regions include, without limitation, promoter sequences, enhancer sequences, response elements, protein recognition sites, inducible elements, protein binding sequences, 5'and 3'untranslated regions (UTR), transcription initiation sites, termination sequences, polyadenylation sequences. And introns. Such vectors can also be used to produce chimeric polypeptides by expression, which are suitable host cells such as CHO cells.

Preferred promoters that control transcription from vectors in mammalian host cells are from a variety of sources, such as polyoma, simian virus 40 (SV40), adenovirus, retrovirus, hepatitis B virus, and most preferably cytomegalovirus ( CMV), or from a heterologous mammalian promoter, such as a β-actin promoter or EF1α promoter, or a hybrid or chimeric promoter (eg, a CMV promoter fused to a β-actin promoter). Of course, promoters from host cells or related species are also useful herein.

Enhancer generally refers to a sequence of DNA that functions at a non-fixed distance from the start site of transcription and can be 5'or 3'to a transcription unit. Moreover, enhancers can be within the introns as well as within the coding sequence itself. They are generally 10 to 300 base pairs (bp) in length and function as cis. Enhancers generally function to increase transcription from nearby promoters. Enhancers may also contain responsive elements that mediate the regulation of transcription. Although many enhancer sequences are known in mammalian genes (globin, elastase, albumin, fetal protein, and insulin), enhancers will typically be used from eukaryotic viruses for general expression. Preferred examples are the SV40 enhancer at the end of the origin of replication, cytomegalovirus early promoter enhancer, the polyoma enhancer at the end of the origin of replication and adenovirus enhancer.

Promoters and/or enhancers can be inducible (eg, chemically or physically regulated). Chemically regulated promoters and/or enhancers can be regulated, for example, by the presence of alcohols, tetracyclines, steroids or metals. Physically regulated promoters and/or enhancers can be regulated by environmental factors such as temperature and light, for example. Optionally, the promoter and/or enhancer region can act as a constitutive promoter and/or enhancer to maximize expression of the transcription unit region to be transcribed. In certain vectors, promoters and/or enhancer regions can be activated in a cell type specific manner. Optionally, the promoter and/or enhancer region in a particular vector may be active in all eukaryotic cells, regardless of cell type. Preferred promoters of this type are CMV promoter, SV40 promoter, β-actin promoter, EF1α promoter and retroviral long terminal repeat (LTR).

Vectors may also contain markers and/or origins of replication, for example. Marker genes can confer to cells a selectable phenotype, for example antibiotic resistance. The marker product is used to determine whether the vector has been delivered to the cell and is expressed when delivered. Examples of selectable markers for mammalian cells are dihydrofolate reductase (DHFR), thymidine kinase, neomycin, neomycin analog G418, hygromycin, puromycin and blasticidin. When these selectable markers are successfully delivered to a mammalian host cell, the transformed mammalian host cell can survive when placed under selective pressure. Examples of other markers include, for example, the E. coli lacZ gene, green fluorescent protein (GFP) and luciferase. In addition, the expression vector may contain a tag sequence designed to facilitate manipulation or detection (eg, purification or localization) of the expressed polypeptide. Tag sequences, such as GFP, glutathione S-transferase (GST), polyhistidine, c-myc, hemagglutinin, or FLAG™ tag (Kodak; New Haven, CT) sequence, are typically associated with the encoded polypeptide. Expressed by fusion. Such tags can be inserted anywhere in the polypeptide, including the carboxyl or amino terminus.

As used herein, the term peptide, polypeptide or protein is used broadly to mean two or more amino acids linked by peptide bonds. Proteins, peptides and polypeptides are also used interchangeably herein to refer to amino acid sequences. It should be appreciated that the term polypeptide is not used herein to suggest a specific size or number of amino acids comprising a molecule and that the peptides of the invention may contain at most several amino acid residues or more. As used entirely, subjects are vertebrates, more specifically mammals (e.g., humans, horses, cats, dogs, cows, pigs, sheep, goats, mice, rabbits, rats and guinea pigs), birds, reptiles, amphibians , Fish and other animals. This term does not refer to a specific age or gender. Thus, it is intended to include adult and neonatal subjects, whether male or female. As used herein, patient or subject can be used interchangeably and can refer to a subject having a disease or disorder (eg, cancer). The term patient or subject includes human and veterinary subjects.

Subjects at risk of developing a disease or disorder may be genetically predisposed to the disease or disorder, for example, have a family history or have a mutation in the gene causing the disease or disorder, or exhibit early signs or symptoms of the disease or disorder. I can. A subject with a current disease or disorder may have one or more symptoms of the disease or disorder and have been diagnosed with the disease or disorder.

The methods and formulations described herein are useful for both prophylactic and therapeutic treatment. For prophylactic use, a therapeutically effective amount of a chimeric polypeptide described herein or a chimeric nucleic acid sequence encoding a chimeric polypeptide may be administered prior to onset (e.g., before obvious signs of cancer or inflammation) or during early onset (e.g. For example, at early signs and symptoms of cancer or inflammation). Prophylactic administration can occur several days to years before symptoms of cancer or inflammation appear. For example, prophylactic administration can be used in prophylactic treatment of a subject diagnosed as having a genetic predisposition to cancer. Therapeutic treatment comprises administering to the subject a therapeutically effective amount of a chimeric polypeptide described herein or a nucleic acid sequence encoding a chimeric polypeptide described herein after diagnosis or development of cancer or inflammation (e.g., an autoimmune disease). . Prophylactic use can also be applied when a patient with anticipated inflammation is undergoing treatment, eg chemotherapy.

According to the methods taught herein, an effective amount of an agent (eg, chimeric polypeptide) is administered to a subject. The terms effective amount and effective dosage are used interchangeably. The term effective amount is defined as any amount necessary to produce the desired physiological response. The effective amount and schedule for administering the formulation can be determined empirically, and it is within the skill of the art to make this determination. The dosage range for administration is a range large enough to produce the desired effect in which one or more symptoms of the disease or disorder are affected (eg, reduced or delayed). Dosage should not be large enough to cause real side effects such as unwanted cross-reaction, anaphylactic reaction, etc. In general, the dosage may vary depending on age, condition, sex, type of disease, degree of disease or disorder, route of administration or whether other drugs are included in the therapy, and may be determined by one of skill in the art. Dosage can be adjusted by individual physicians if there are any contraindications. The dosage may vary and may be administered in one or more doses daily for a day or several days. Guidance on the appropriate dosage for a given class of medicinal products can be found in the literature.

The term treating, treating or treating, as used herein, refers to a method of reducing the effect of a disease or condition or symptoms of a disease or condition. Thus, treatment in the disclosed methods is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% at the severity of the established disease or condition or at the symptoms of the disease or condition. May refer to reduction. For example, a method of treating a disease is considered treatment if there is a 10% reduction in one or more symptoms of the disease in a subject compared to a control. Thus, the reduction is a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100% or any percentage reduction between 10% and 100% compared to the congenital or control level. Can be It is understood that treatment does not necessarily refer to the cure or complete resection of a disease, condition, or symptom of a disease or condition.

As used herein, the terms prevent, prevent and prevent a disease or disorder, for example, inhibiting or delaying the onset or exacerbation of one or more symptoms of the disease or disorder, wherein the subject is one of the disease or disorder It refers to an action, which is the administration of a chimeric polypeptide or a nucleic acid sequence encoding a chimeric polypeptide that occurs before or approximately simultaneously with the time on which the above symptoms begin to appear. As used herein, reference to alleviation, reduction or inhibition is a change of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more compared to the control level. Includes. These terms may include complete removal, but not necessarily.

IL-2 variants that are selective for IL2Rαβγ compared to IL2Rβγ have been developed (Shanafelt, AB, et al., 2000, Nat Biotechnol. 18:1197-202; Cassell, DJ, et al., 2002, Curr Pharm Des., 8:2171- 83). These variants have amino acid substitutions that reduce their affinity for IL2Rβ. Since IL-2 has an undetectable affinity for IL2Rγ, these variants consequently have a reduced affinity for the IL2Rβγ receptor complex and a reduced ability to activate IL2Rβγ-expressing cells, but the ability to bind to IL2Rα and IL2Rαβγ Retains the ability to bind and activate receptor complexes.

One of these variants, IL2/N88R (Bay 50-4798), has been clinically tested as a low-toxic version of IL-2 as an immune system stimulant based on the hypothesis that IL2Rβγ-expressing NK cells are a major contributor to toxicity. Bay 50-4798 has been shown to selectively stimulate the proliferation of activated T cells compared to NK cells, and cancer patients (Margolin, K., et al., 2007, Clin Cancer Res., 13:3312-9) and HIV patients (Davey, RT, et al., 2008, J Interferon Cytokine Res., 28:89-100) in Phase I/II clinical trials. These clinical trials indicated that Bay 50-4798 could be much safer and more resistant than aldesleukin, and also indicated that it increased the level of CD4+CD25+ T cells, a cell population rich in Treg cells. Following these tests, studies in the field more fully established the identity of Treg cells and demonstrated that Treg cells selectively express IL2Rαβγ (reviewed in Malek, TR, et al., 2010, Immunity, 33:153-65. being).

In addition, mutants that selectively alter the affinity for the CD25 chain compared to native IL-2 can be prepared.

IL-2 is generally different from that of the IL-2R complex or wild-type IL-2 corresponding to the IL-2Rα subunit or of the currently available mutants (referred to as C125S, since the cysteine residue is replaced by a serine residue at position 125). It can be engineered to produce mutants that specifically bind with affinity.

Accordingly, the present invention provides a mutant interleukin-2 (IL-2) comprising an amino acid sequence that is at least 80% identical (e.g., 85, 87, 90, 95, 97, 98, or 99% identical) to wild-type IL-2. ^* ) Characterize the polypeptide and binds higher to the IL-2 trimer receptor compared to the dimeric IL-2 receptor compared to WT IL-2. Typically, the mutein will also bind to the IL-2 receptor α subunit (IL-2Rα) with an affinity greater than that of wild type IL-2 to IL-2Rα. The amino acid sequence in the mutant IL-2 polypeptide may differ from SEQ ID NO: 1 (UniProtKB accession number P60568) because it contains (or only contains) one or more amino acid substitutions that can be considered conservative or non-conservative substitutions. have. Non-naturally occurring amino acids may also be included. Alternatively or additionally, the amino acid sequence may differ from SEQ ID NO: 1 (which may be considered a “reference” sequence) due to the inclusion and addition and/or deletion of one or more amino acid residues. More specifically, the amino acid sequence may differ from that of SEQ ID NO: 1 due to mutations in at least one of the following positions of SEQ ID NO: 1: 1, 4, 8, 9, 10, 11, 13, 15, 26, 29 , 30, 31, 35, 37, 46, 48, 49, 54, 61, 64, 67, 68, 69, 71, 73, 74, 75, 76, 79, 88, 89, 90, 92, 99, 101 , 103, 114, 125, 128, or 133 (or combinations thereof). As mentioned, it can be 2, 3, 4, 5, 6, 7, 8, 9, 10 or 11 or more of the positions (including at most all), but these positions At least one of them may be changed. For example, the amino acid sequence may differ from SEQ ID NO: 1 at positions 69 and 74 and further at one or more of positions 30, 35 and 128. The amino acid sequence may also differ from SEQ ID NO: 2 (as disclosed in US 7569215, incorporated herein by reference) in one of the following sets of positions: (a) positions 64, 69 and 74; (b) positions 69, 74 and 101; (c) positions 69, 74 and 128; (d) positions 30, 69, 74 and 103; (e) positions 49, 69, 73 and 76; (f) positions 69, 74, 101 and 133; (g) positions 30, 69, 74 and 128; (h) positions 69, 74, 88 and 99; (i) positions 30, 69, 74 and 128; (j) positions 9, 11, 35, 69 and 74; (k) positions 1, 46, 49, 61, 69 and 79; (l) positions 48, 68, 71, 90, 103 and 114; (m) positions 4, 10, 11, 69, 74, 88 and 133; (n) positions 15, 30 31, 35, 48, 69, 74 and 92; (O) positions 30, 68, 69, 71, 74, 75, 76 and 90; (p) positions 30, 31, 37, 69, 73, 74, 79 and 128; (q) positions 26, 29, 30, 54, 67, 69, 74 and 92; (r) positions 8, 13, 26, 30, 35, 37, 69, 74 and 92; And (s) positions 29, 31, 35, 37, 48, 69, 71, 74, 88 and 89. In addition to the mutations at these positions, the amino acid sequence of the mutant IL-2 polypeptide is otherwise with SEQ ID NO: 1 It can be the same. With regard to certain substitutions, the amino acid sequence may differ from SEQ ID NO: 1 because it has one or more of the following mutations: A1T, S4P, K8R, K9T, T10A, Q11R, Q13R, E15K, N26D, N29S, N30S, N30D, N30T, Y31H, Y31C, K35R, T37A, T37R, M46L, K48E, K49R, K49E, K54R, E61D, K64R, E67G, E68D, V69A, N71T, N71A, N71R, A73V, Q74P, S75P, K76E, K76E H79R, N88D, I89V, N90H, I92T, S99P, T101A, F103S, I114V, I128T, I128A, T133A, or T133N. Our nomenclature is consistent with the nomenclature of the scientific literature, wherein the single letter code of the amino acid in the wild-type or reference sequence is followed by its position in the sequence followed by the single letter code of the amino acid in which the amino acid is replaced. Thus, A1T designates the substitution of alanine residue position 1 for threonine. Other mutant polypeptides within the scope of the present invention include those comprising the mutant of SEQ ID NO: 2 with substitutions at V69 (eg A) and Q74 (eg P). For example, the amino acid sequence may comprise one of the following sets of mutations for SEQ ID NO: 2: (a) K64R, V69A, and Q74P; (b) V69A, Q74P, and T101A; (c) V69A, Q74P, and I128T; (d) N30D, V69A, Q74P, and F103S; (e) K49E, V69A, A73V, and K76E; (f) V69A, Q74P, T101A, and T133N; (g) N30S, V69A, Q74P, and I128A; (h) V69A, Q74P, N88D, and S99P; (i) N30S, V69A, Q74P, and I128T; (j) K9T, Q11R, K35R, V69A, and Q74P; (k) A1T, M46L, K49R, E61D, V69A, and H79R; (l) K48E, E68D, N71T, N90H, F103S, and I114V; (m) S4P, T10A, Q11R, V69A, Q74P, N88D, and T133A; (n) E15K, N30S Y31H, K35R, K48E, V69A, Q74P, and I92T; (o) N30S, E68D, V69A, N71A, Q74P, S75P, K76R, and N90H; (p) N30S, Y31C, T37A, V69A, A73V, Q74P, H79R, and I128T; (q) N26D, N29S, N30S, K54R, E67G, V69A, Q74P, and I92T; (r) K8R, Q13R, N26D, N30T, K35R, T37R, V69A, Q74P, and I92T; And (s) N29S, Y31H, K35R, T37A, K48E, V69A, N71R, Q74P, N88D, and I89V. SEQ ID NO: 2 is disclosed in US 7569215, which is incorporated herein by reference as an exemplary IL-2 polypeptide sequence that can be used in the present invention.

As mentioned above, any mutant IL-2 polypeptide disclosed herein may comprise the described sequence; They may also be limited to the described sequence, otherwise they are identical to SEQ ID NO: 1. Moreover, any of the mutant IL-2 polypeptides described herein may optionally comprise a substitution of a cysteine residue at position 125 for another residue (e.g., serine) and/or the position of SEQ ID NO: 1. It may optionally include a deletion of an alanine residue in 1.

_{The mutant IL-2 polypeptide disclosed herein has a K d} of less than about 28 nM (eg, less than about 25 nM; less than about 5 nM; about 1 nM; less than about 500 pM; or less than about 100 pM). Can bind to the IL-2Rα subunit. More specifically, the mutant IL-2 polypeptide may have an affinity equilibrium constant of less than 1.0 nM (eg, about 0.8, 0.6, 0.4 or 0.2 nM). Affinity can also be expressed as the relative rate of dissociation from the IL-2Rα subunit or IL-2 receptor complex (eg, a complex expressed on the surface of a cell or otherwise a membrane bound complex). For example, mutant IL-2 polypeptides can dissociate at a reduced rate compared to wild-type polypeptides or IL-2 based therapeutics such as IL-2 ^{*, for example from IL-2Rα.} Alternatively, affinity ^{can be characterized by the duration or average time the IL-2*} polypeptide lasts on the surface of, for example, a cell expressing IL-2R. For example, the IL-2 ^* polypeptide can persist at the receptor for at least about 2, 5, 10, 50, 100 or 250 hours (or more).

Materials, compositions and components that can be used in, can be used in conjunction with, are used in the preparation, or are products of the disclosed methods and compositions are disclosed. When these and other substances are disclosed herein, and combinations, subsets, interactions, groups, etc. of these substances are disclosed, specific references to each of the various individual and collective combinations and permutations of these compounds are expressly disclosed. It is understood that it may not be, and each is specifically contemplated and described herein. For example, if a method is disclosed and discussed and a number of modifications that can be made to a number of molecules, including the method, are discussed, each and all combinations and permutations of the method, and possible modifications, are not specifically indicated to the contrary. It is specifically considered. Likewise, any subset or combination of these is also specifically contemplated and disclosed. This concept applies to all aspects of the present disclosure, including, but not limited to, steps of a method of using the disclosed compositions. Thus, where there are various additional steps that can be performed, each of these additional steps can be performed with any particular method step or combination of method steps of the disclosed method, and each such combination or subset of combinations is specifically It is understood that it should be considered and considered public.

The publications cited herein and the materials to which they are cited are specifically incorporated herein by reference in their entirety.

Example

The following are examples of the methods and compositions of the present invention. It is understood that various other embodiments may be practiced given the general description provided herein.

Example 1: Detection of IL-2, IL-2 mutein, IL-2Rα and IL-2Rγ in fusion proteins by ELISA

The IL-2 mutein is detected with commercially available antibodies, such as anti-IL-2 monoclonal (JES6-1A12) (BD Pharmingen; San Jose, CA). Positive controls are used to indicate whether monoclonal antibodies recognize cytokines or muteins. Antibodies against the IL-2Rα and IL-2Rγ chains are also used. Wells of the 96-well plate are coated with antibody (2.5 μg/ml) in PBS. Block the wells with 5% fat-free milk in PBS with 0.2% Tween®20 (PBS-M-Tw) and add the fusion protein at 37° C. for 1-2 hours. After washing, an anti-IL-2 biotin-labeled antibody such as JES5H4 (BD Pharmingen) is added and binding is detected using Strepavidin HRP (Southern Biotechnology Associates; Birmingham, Alabama). ELISA plates were developed by adding 50 μl O-phenylenediamine (OPD) (Sigma-Aldrich) to 0.1 M citrate pH 4.5 and 0.04% H ₂ O ₂ , and stopped by adding _{50 μl/well 2N H 2} SO ₄ The absorbance was read at 490 nm.

Example 2: Protease cleavage of fusion proteins by MMP9 protease

Those of skill in the art will be familiar with how to set up protein cleavage assays. 100 ug of protein in 1xPBS pH 7.4 was digested with 1 ug of active MMP9 (Sigma catalog number SAE0078-50 or Enzo catalog BML-SE360) and incubated at room temperature for up to 16 hours. The isolated protein is continuously used for functional analysis or stored at -80°C prior to testing. The extent of cleavage was monitored by SDS PAGE using methods well known in the art. As shown in Fig. 10 , complete cleavage of the fusion protein by the MMP9 protease is observed.

Example 3: CTLL-2 analysis

CTLL2 cells (ATCC) were plated in suspension at a concentration of 500,000 cells/well in culture medium with or without 40 mg/ml human serum albumin (HSA), and serial recombinant hIL2 at _{37° C. and 5% CO 2 for 72 hours.} Or stimulated with activatable hIL2 dilution. The activity of uncleaved and cleaved activatable hIL2 was tested. The cleaved activatable hIL2 was produced by incubation with active MMP9. Cellular activity was assessed using CellTiter-Glo (Promega) luminescence-based cell viability assay. The results are shown in Figures 7-9 .

Example 4: Protease cleavage of IL-2/IL-2Rα/IL-2Rγ chimeric polypeptide results in increased accessibility to antibodies and biologically active IL-2 muteins

The IL-2 mutein fusion protein is characterized biochemically before and after cleavage with a protease, such as PSA. Immunoblot analysis will show that the fusion protein can be cleaved by PSA and the strength of the predicted low molecular weight cleavage product of approximately 20 kDa reactive with anti-IL-2 antibody increases after processing the sample with PSA. The degree of cleavage depends on the amount of PSA as well as the time of incubation. Interestingly, when the fusion protein was analyzed before and after PSA treatment by ELISA, it was found that the apparent amount of IL-2 increased after PSA cleavage. In this experiment, there is an approximately 2 or 4-fold increase in the apparent amount of IL-2 detected using this sandwich ELISA depending on the construct, which is partially hindered by the antibody binding in the intact fusion protein. Suggests. Aliquots of the same sample can also be analyzed after PSA treatment using a CTLL-2 cell line that requires IL-2 for growth and survival, and colorimetric MTT assays can be used to confirm the viability of cells. In this assay, the more the supernatant is diluted, the more biologically active IL-2 is contained and the amount of biologically active IL-2 increases after PSA cleavage. The amount of IL-2 mutein increase is determined by the increase in predicted low molecular weight cleavage fragment of approximately 20 kDa reactive with anti-IL-2 antibody after PSA cleavage, increase in antibody accessibility, and most importantly, biologically active IL. -2 will suggest that there is an increase in the amount of muteins.

Example 5. Protease activated fusion protein In vivo Delivery results in reduced tumor growth

Chimeric polypeptides are tested to see if they can have biological effects in vivo. For these experiments, a system in which tumor cells injected intraperitoneally adhere rapidly and preferentially to milky spots, a series of organized immune aggregates found in the serous membrane, and grow early is used (Gerber et al., Am. J. Pathol). 169:1739-52 (2006)). This system provides a convenient way to examine the effect of fusion protein therapy on tumor growth because the fusion protein can be delivered multiple times intraperitoneally and the cells of the dissociated serous membrane can be examined to analyze tumor growth. For these experiments, the colon 38 cell line, a fast growing tumor cell line that expresses both MMP2 and MMP9 in vitro, can be used. The serous tissue generally expresses relatively small amounts of MMP2 and MMP9, but the presence of colon 38 tumors on the serous increases MMP levels. Using this tumor model, the ability of the IL-2 mutein fusion protein to influence tumor growth is investigated. Colon 38 cells are injected intraperitoneally to adhere and grow for 1 day and then intraperitoneally treated with fusion protein daily. On day 7, animals are sacrificed and the serous membrane is examined for tumor growth using flow cytometry and colony-forming assays.

Example 6: Construction of an exemplary activatable IL2 protein targeting CD20

Generation of activatable IL2 domain

IL-2 polypeptides capable of binding to CD20 polypeptides present in tumors or tumor cells are produced as follows. A nucleic acid is generated that (1) encodes an IL-2 polypeptide sequence and (2) contains one or more polypeptide linkers: the nucleic acid sequence. The activatable interleukin plasmid construct may have an optional Flag, His or other affinity tag and is electroporated and purified into HEK293 or other suitable human or mammalian cell line. Validation assays include T cell activation assays using T cells responsive to IL-2 stimulation in the presence of a protease.

Generation of scFv CD20 binding domain

CD20 is one of the cell surface proteins present in B-lymphocytes. The CD20 antigen is found in normal and malignant pre-B and mature B lymphocytes, including those present in more than 90% of B-cell non-Hodgkin's lymphoma (NHL). The antigen is absent from hematopoietic stem cells, activated B lymphocytes (plasma cells) and normal tissues. As such, several antibodies of primarily murine origin have been described: 1F5, 2B8/C2B8, 2H7 and 1H4.

Thus, human or humanized anti-CD20 antibodies are used to generate scFv sequences for the CD20 binding domain of the activatable interleukin protein. DNA sequences encoding human or humanized VL and VH domains are obtained and codons for constructs are optionally optimized for expression in cells from Homo sapiens. The order in which the VL and VH domains appear in the scFv varies (ie, in the VL-VH or VH-VL direction), and the “G4S” or “G ₄ S” subunit (G ₄ S) ₃ connects the variable domains to the scFv domain Create Anti-CD20 scFv plasmid constructs can have an optional Flag, His or other affinity tag and are electroporated and purified into HEK293 or other suitable human or mammalian cell lines. Validation assays include binding assays by FACS, kinetic assays using Proteon and staining of CD20-expressing cells.

Cloning of DNA expression constructs encoding activatable IL2 protein

The activatable IL2 construct with a protease cleavage site domain is used in combination with an anti-CD20 scFv domain and a serum half-life extending element (e.g., HSA binding peptide or VH domain) to construct an activatable interleukin protein. For expression of activatable interleukin proteins in CHO cells, the coding sequences of all protein domains are cloned into a mammalian expression vector system. Briefly, gene sequences encoding activatable interleukin domains, serum half-life extending elements, and CD20 binding domains together with peptide linkers L1 and L2 are synthesized and subcloned separately. The resulting constructs were then ligated together in the order of CD20 binding domain-L1-IL2 subunit 1-L2-protease cleavage domain-L3-IL2 subunit 2-L4-anti-CD20 scFv-L5-serum half-life extending element. Create the final construct. All expression constructs are designed to contain coding sequences for the N-terminal signal peptide and C-terminal hexahistidine (6xHis)-tag, respectively, to facilitate protein secretion and purification.

Expression of IL2 protein that can be activated in stably transfected CHO cells

CHO cell expression system (Flp-In®, Life Technologies), a derivative of CHO-K1 Chinese hamster ovary cells (ATCC, CCL-61) (Kao and Puck, Proc. Natl. Acad Sci USA 1968;60(4):1275 -81) is used. Adherent cells are passaged according to standard cell culture protocols provided by Life Technologies.

For adaptation to growth in suspension, cells are separated from tissue culture flasks and placed in serum-free medium. Suspension-adapted cells are cryopreserved in medium with 10% DMSO.

Recombinant CHO cell lines stably expressing the secreted activatable interleukin protein are produced by transfection of suspension-adapted cells. During selection with the antibiotic hygromycin B, the viable cell density is measured twice a week, the cells are centrifuged and resuspended in fresh selection medium to a maximum density of ^{0.1x10 6 viable cells/mL.} The cell pool stably expressing the activatable interleukin protein is recovered after 2-3 weeks of selection at the time the cells are transferred from the shake flask to the standard culture medium. Expression of the recombinant secreted protein is confirmed by performing protein gel electrophoresis or flow cytometry. Stable cell pools are cryopreserved in DMSO containing medium.

The activatable IL2 protein is produced in 10-day fed-batch culture of stably transfected CHO cell lines by secretion into the cell culture supernatant. Cell culture supernatant is typically harvested after 10 days at a culture viability of >75%. Every other day, samples are collected from production cultures and cell density and viability are assessed. On harvest day the cell culture supernatant is cleared by centrifugation and vacuum filtration prior to further use.

Protein expression titer and product integrity in cell culture supernatant are analyzed by SDS-PAGE.

Purification of activatable IL2 protein

The activatable IL2 protein is purified from the CHO cell culture supernatant in a two-step procedure. The construct is subjected to affinity chromatography in the first step and then preparative size exclusion chromatography (SEC) for Superdex 200 in the second step. Samples are buffer-exchanged and concentrated by ultrafiltration to a typical concentration of >1 mg/mL. The purity and homogeneity (typically >90%) of the final sample was assessed by SDS PAGE under reducing and non-reducing conditions, followed by immunoblotting with anti-HSA or anti-idiotype antibodies, respectively, as well as analytical SEC. . Purified protein is stored as aliquots at -80°C until use.

Example 7: Determination of antigen affinity by flow cytometry

The activatable IL2 protein of Example 6 is tested for binding affinity to ^{human CD20 +} cells and cynomolgus CD20 ^{+ cells.}

CD20 ⁺ cells are incubated with 100 μL serial dilutions of the activatable interleukin protein of Example 6 and at least one protease. After washing three times with FACS buffer, cells are incubated with 0.1 mL of 10 μg/mL mouse monoclonal anti-idiotype antibody in the same buffer for 45 minutes on ice. After the second wash cycle, cells are incubated with 0.1 mL of 15 μg/mL FITC-conjugated goat anti-mouse IgG antibody under the same conditions as before. As a control, cells are incubated with anti-His IgG followed by FITC-conjugated goat anti-mouse IgG antibody without activatable IL2 protein. The cells were then washed again and resuspended in 0.2 mL of FACS buffer containing 2 μg/mL propidium iodide (PI) to remove dead cells. Fluorescence of 1x10 ⁴ living cells was measured using a Beckman-Coulter FC500 MPL flow cytometer (Beckman-Coulter, Krefeld, Germany) using MXP software, or a Millipore Guava EasyCyte flow cytometer (Merck Millipore, Schwalbach, Germany) using Incyte software. Is measured using. The average fluorescence intensity of the cell sample is calculated using CXP software (Beckman-Coulter, Krefeld, Germany) or Incyte software (Merck Millipore, Schwalbach, Germany). After subtracting the fluorescence intensity values of cells stained with only the secondary and tertiary reagents, the value is an equation for single-site binding (hyperbolic) of GraphPad Prism (version 6.00 for Window, GraphPad Software, La Hora, CA). It is used to calculate the _{K D value.}

CD20 binding and cross-reactivity are ^{evaluated in human CD20 +} tumor cell lines. _{The K D} ratio of cross-reactivity is calculated using the _{K D} value determined in a CHO cell line expressing a recombinant human or recombinant cynomolgus antigen.

Example 8: Cytotoxicity assay

The activatable IL2 protein of Example 6 is evaluated in vitro for mediating an immune response against ^{CD20 + target cells.}

Fluorescently labeled CD20 ⁺ REC-1 cells (mantle cell lymphoma cell line, ATCC CRL-3004) were the activatable of Example 5 with CB15 T-cells (standardized T-cell line) as isolated PBMCs or effector cells of random donors. It is incubated in the presence of the IL2 protein and one or more proteases. After incubation at 37° C. for 4 hours in a humidified incubator, fluorescence dye emission from the target cells to the supernatant is measured in a spectrofluorimeter. The target cells incubated without the activatable IL2 protein of Example 1 and the target cells completely lysed by the addition of saponin at the end of the incubation served as negative and positive controls, respectively.

Based on the measured remaining live target cells, the specific cell lysis percentage is calculated according to the following formula: [1-(number of live targets _(sample) /number of live targets _(voluntary) )] x 100%. Sigmoidal dose response curves and EC ₅₀ values are calculated by non-linear regression/4-parameter logistic fit using GraphPad software. The dissolution values obtained for a given antibody concentration are used to calculate sigmoidal dose-response curves by a four parameter logistic fit assay using Prism software.

Example 9: Pharmacokinetics of activatable IL2 protein

The activatable IL2 protein of Example 5 is evaluated for half-life elimination in animal studies.

The activatable IL2 protein is administered to cynomolgus monkeys by 0.5 mg/kg bolus injection into the saphenous vein. Another group of cynomolgus monkeys received similarly sized IL2 constructs but lacked a serum half-life prolonging factor. Groups 3 and 4 were each administered an IL2 construct with a serum half-life extending element and a cytokine with CD20 and a serum half-life extending element, both of which are similar in size to the activatable interleukin protein. Each test group consists of 5 monkeys. Serum samples are taken at designated time points and serially diluted, and the concentration of protein is determined using a binding ELISA to CD20.

Pharmacokinetic analysis is performed using the test item plasma concentration. The group mean plasma data for each test item follows a multi-index profile when plotted against post-dose time. The data fit a standard two-compartment model with bolus influx and first order rate constants for the distribution and removal steps. The general equation for optimal fit of the data for iv administration is: c(t) = Ae ^-αt + Be- ^βt , where c(t) is the plasma concentration at time t, and A and B are Y- Is the intercept of the axis, α and β are the apparent first order rate constants for the distribution and removal steps, respectively. The α-stage is the initial stage of clearance and reflects the distribution of the protein into all extracellular fluids of the animal, while the second or β-stage portion of the decay curve represents true plasma clearance. How to fit these equations is well known in the art. For example, A = D/V(α-k21)/(α-β), B = D/V(β-k21)/(α-β), α and β (if α>β) are secondary Equation: r ² +(k12+k21+k10)r+k21k10=root of 0 and V = volume of distribution, k10 = removal rate, k12 = rate of transfer from section 1 to section 2, and k21 = transfer from section 2 to section 1 Rate, and D = estimated parameters of the administered dose are used.

Data Analysis: A graph of the concentration versus time profile was created using KaleidaGraph (KaleidaGraph™ V. 3.09 Copyright 1986-1997. Synergy Software. Reading, PA). Values reported below reportable (LTR) are not included in the PK analysis and are not displayed graphically. Pharmacokinetic parameters are determined by compartment analysis using WinNonlin software (WinNonlin® Professional V. 3.1 WinNonlin™ Copyright 1998-1999. Pharsight Corporation, Mountain View, CA). Pharmacokinetic parameters are calculated as described in Ritschel WA and Kearns GL, 1999, IN: Handbook Of Basic Pharmacokinetics Including Clinical Applications, 5th edition, American Pharmaceutical Assoc., Washington, DC.

The activatable IL2 protein of Example 5 is expected to have improved pharmacokinetic parameters such as an increase in elimination half-life compared to a protein that lacks a serum half-life extending element.

Example 10: Xenograft tumor model

The activatable IL2 protein of Example 5 is evaluated in a xenograft model.

Female immune-deficient NOD/scid mice are lethal-down irradiated (2 Gy) and inoculated subcutaneously with ^{4x10 6 Ramos RA1 cells on the right dorsal flank.} When tumors ^{reach 100-200 mm 3} animals are assigned to 3 treatment groups. Groups 2 and 3 (8 animals each) ^{injected 1.5×10 7} activated human T-cells intraperitoneally. After 3 days, animals in Group 3 were subsequently treated with a total of 9 intravenous doses (qdx9d) of 50 μg activatable interleukin protein of Example 1. Groups 1 and 2 are treated as vehicles only. Body weight and tumor volume are measured over 30 days.

Animals treated with the activatable IL2 protein of Example 5 are expected to have a statistically significant delay in tumor growth compared to each vehicle-treated control.

While preferred embodiments of the present invention have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. Various modifications, changes and substitutions will now occur to those skilled in the art without departing from the present invention. It should be understood that various alternatives to the embodiments of the invention described herein can be used to practice the invention. The following claims define the scope of the invention, and it is intended that methods and structures within the scope of these claims and their equivalents are covered thereby.

Example 11: HEK Blue analysis

HEK-Blue IL2 cells (InvivoGen) were plated in suspension at a concentration of 50,000 cells/well in culture medium with or without 15-40 mg/ml human serum albumin (HSA) and with serial recombinant hIL2 or activatable hIL2 dilutions. Stimulated at 37° C. and 5% CO _{2 for 24 hours.} The activity of uncleaved and cleaved activatable hIL2 was tested. The truncated inducible hIL2 was generated by incubation with active MMP9. IL12 activity was evaluated by quantifying the secreted alkaline phosphatase (SEAP) activity using the reagent QUANTI-Blue (InvivoGen) as a colorimetric assay. The results are shown in FIG. 11 .

Example 12: MC38 experiment

The MC38 cell line, a fast growing colon adenocarcinoma cell line expressing MMP9 in vitro, was used. Using this tumor model, the ability of the fusion protein to influence tumor growth was investigated.

Example 12a MC38 IL- 2POC

Formulation and treatment:

^# -Control

Example 12b: MC38 IL-2

Formulation and treatment:

^# -Control

Example 12c: Treatment with ACP16, ACP132 and ACP21

Mice were anesthetized with isoflurane for cell transplantation to reduce ulcers. CR female C57BL/6 mice were set flank with ⁵ ×10 5 MC38 tumor cells in 0% Matrigel sc. The cell injection volume was 0.1 mL/mouse. The age of the mice on the day of initiation was 8-12 weeks old. Pair matching was performed when tumors reached an average size of 100-150 mm ^{3 and started treatment.} Body weight was measured initially and then every other week to the end. Caliper measurements were carried out every other week to the end. Any adverse reactions were reported immediately. All individual animals with a single observation of 3 consecutive measurements of >30% weight loss or >25% weight loss were euthanized. Any group with an average weight loss of >20% or >10% mortality discontinued dosing: the group was not euthanized and recovery was allowed. Individuals who reached individual weight loss endpoints within groups with >20% weight loss were euthanized. When the group treatment-related weight loss recovered to within 10% of the original body weight, dosing was resumed with a lower dose or less frequent dosing schedule. An exception to non-treatment% body weight recovery was allowed on a case-by-case basis. The endpoint was tumor growth delay (TGD). Animals were monitored individually. The endpoint of the experiment was ^{a tumor volume of 1500 mm 3} or 45 days, whichever came first. Respondents were tracked longer. Upon reaching the end point, the animal should be euthanized. The results are shown in Figure 17.

Example 12d: MC38 re-attack

Mice cured from Example 12b (ACP16-treated) were re-attacked with tumor implantation to determine whether anti-tumor memory was established from initial treatment.

Formulation and treatment:

^# -Control

step:

Mice were anesthetized with isoflurane for cell transplantation to reduce ulcers. This part of the study began on the day of transplantation (Day 1). Group 1 consisted of 33 CR female C57BL/6 mice set with ⁵ ×10 5 MC38 tumor cells in 0% Matrigel subcutaneously on the flank. Groups 2-6 consisted of 33 CR female C57BL/6 mice set with ⁵ ×10 5 MC38 tumor cells in 0% Matrigel sc on the left flank. Tumors from the previous MC38 experiment (Example 12b) were implanted in the right flank of each animal. The cell injection volume was 0.1 mL/mouse. Initially, the age of control mice was 14 to 17 weeks old. These mice were age matched to mice from the previous MC38 experiment (Example 12b). During the re-attack, no dose of the active agent occurred. The body weight was measured every other week until the end, similar to the caliper measurement. Any adverse reactions or deaths were reported immediately. All individual animals with a single observation of 3 consecutive measurements of >30% weight loss or >25% weight loss were euthanized. The endpoint was tumor growth delay (TGD). Animals were monitored individually. The endpoint of the experiment was ^{a tumor volume of 1000 mm 3} or 45 days, whichever came first. Respondents were tracked longer if possible. Animals were euthanized upon reaching the endpoint. The results are shown in Figure 15.

Example 13. Conditionally active fusion protein containing a blocking moiety that is a serum albumin binding domain

This example is a fusion protein, preferably, having inducible activity, i.e., they are typically inactive until induced by separation of the blocking moiety from the active moiety upon cleavage of the linker between the blocking moiety and the active moiety. The following describes the production and activity of cytokines. The fusion protein contains a single antibody variable domain (dAb) that binds serum albumin through a CDR loop and binds to an active moiety (here anti-CD3 scFV) through one or more non-CDR loops (e.g., C loop). do. The serum albumin-binding blocking moiety is operably linked to the active moiety through a protease cleavable linker, and the active moiety is a target domain (here anti-epidermal growth factor receptor (EGFR) dAb or Anti-prostate-specific membrane antigen (PSMA) dAb)). These fusion proteins can be administered as inactive proteins that are activated upon cleavage of the protease cleavable linker and subsequent release of the inhibitory albumin-binding domain. The anti-CD3 scFV in the fusion protein is a surrogate for the desired cytokine in the fusion protein described herein. Containing an albumin-binding dAb that binds and inhibits a desired cytokine (e.g., IL-2, IL-12, interferon) or a functional fragment or mutein thereof, a targeting domain and a cytokine or a functional fragment or mutein thereof Similar fusion proteins can be made using the methods described and illustrated herein. Anti-serum albumin dAb, which binds and inhibits the activity of a desired cytokine or functional fragment or mutein thereof (via cytokine proximity to bound serum albumin) and steric masking of cytokines (non-CDR loops (e.g. For example, it is possible to provide both specific masking of cytokines) through binding to cytokines through C loop). Anti-serum albumin dAbs that bind and inhibit the activity of a desired cytokine or functional fragment or mutein thereof are used to introduce amino acid sequence diversity into the non-CDR loop (e.g., C loop) of the anti-serum albumin binding dAb. It can be obtained using any suitable method such as screening for binding to the desired cytokine. Any suitable method can be used for the selection, such as phage display. For example, an exemplary anti-serum albumin dab that can be used goes to the following sequence, the amino acid sequence of the C loop (bold underlined) can be diversified (e.g., randomized) and the resulting dAbs Serum albumin through and through non-CDR loop interactions can be screened for binding to cytokines. If desired, the amino acid sequence of a known cytokine binding peptide can be grafted into the C loop.

EVQLVESGGGLVQPGNSLRLSCAASGFTFSKFGMSWVRQ GGGGGLDGNEEPGG LEWVSSISGSGRDTLYADSVKGRFTISRDNAKTTLYLQMNSLRPEDTAVYYCTIGGSLSVSSQGTLVTVSS (SEQ ID NO: 137)

A. ProTriTAC protease activation In the laboratory Induces remarkably improved activity in.

Recombinant active drug fragment mimicking purified ProTriTAC (prodrug), non-cleavable ProTriTAC [prodrug (non-cleavable)] and protease-activated ProTriTAC (active drug) were combined with recombinant human CD3 in ELISA analysis, validation of flow cytometry Was tested for functional efficacy in binding to human primary T cells purified in, and cytotoxicity validation of T cell-dependent cells.

For ELISA, soluble ProTriTAC protein at the indicated concentration was incubated with immobilized recombinant human CD3e (R&D Systems) in PBS supplemented with 15 mg/ml human serum albumin at room temperature for 1 hour. Plates were blocked using SuperBlock (Thermo Fisher), washed with PBS with 0.05% Tween-20, non-competitive anti-CD3 idiotype monoclonal antibody 11D3 followed by peroxidase-labeled secondary antibody And TMB-ELISA substrate solution (Thermo Fisher).

For flow cytometry, indicated concentrations of soluble ProTriTAC protein were incubated with purified human primary T cells at 4° C. for 1 hour in the presence of PBS with 2% fetal bovine serum and 15 mg/ml human serum albumin. Plates were washed with PBS with 2% fetal bovine serum and detected using AlexaFluor 647-labeled non-competitive anti-CD3 idiotype monoclonal antibody 11D3 and data analyzed using FlowJo 10 (FlowJo, LLC). .

For functional efficacy in the verification of cytotoxicity of T cell-dependent cells, soluble ProTriTAC protein at the indicated concentrations was purified in a 10:1 effector:target cell ratio of resting human T cells (effector cells) and HCT116 cancer cells (target cells). Incubated at 37° C. for 48 hours. The HCT116 target cell line is stably transfected with a luciferase reporter gene, allowing the measurement of specific T cell-mediated cell death by ONE-Glo (Promega).

B. ProTriTAC shows potent protease dependent anti-tumor activity in rodent tumor xenograft models.

ProTriTAC was evaluated for its anti-tumor activity in vivo in HCT116 subcutaneous xenograft tumors mixed with expanded human T cells in immunocompromised NCG mice. Specifically, 5x106 HCT116 cells were mixed with 2.5x106 expanded T cells per mouse on day 0. Take ProTriTAC the next day q.d. It was performed starting via intraperitoneal injection on a x 10 schedule. Tumor volume measurements were determined using caliper measurements and calculated using the formula V = (length x width x width)/2 at the specified time.

C. Expression, purification and stability of exemplary ProTriTAC trispecific molecules

Protein production

The sequence encoding the inducible fusion protein molecule was cloned into the mammalian expression vector pcDNA 3.4 (Invitrogen) preceded by a leader sequence and followed by a 6x histidine tag (SEQ ID NO: 136). Expi293F cells (Life Technologies A14527) were maintained in suspension in Optimum Growth Flasks (Thomson) between 0.2 and 8 x 1e6 cells/ml in Expi 293 medium. Purified plasmid DNA was transfected into Expi293 cells according to the Expi293 Expression System Kit (Life Technologies, A14635) protocol and maintained for 4-6 days after transfection. Alternatively, the sequence encoding the fusion protein molecule was cloned into the mammalian expression vector pDEF38 (CMC ICOS) transfected with CHO-DG44 dhfr-cells, a stable pool was generated, and cultured in production medium for up to 12 days prior to purification. I did. The amount of exemplary fusion protein in conditioned medium was quantified using an Octet RED 96 instrument with Protein A tips (ForteBio/Pall) using a control fusion protein against a standard curve. The conditioned medium from either host cell was filtered and partially purified by affinity and desalting chromatography. The fusion protein was then polished by ion exchange and by fraction pooling formulated in a neutral buffer containing excipient. Final purity is SDS-PAGE and analytical SEC and Chemstation CDS software (Agilent) using an Acquity BEH SEC 200 1.7u 4.6 x 150 mm column (Waters Corporation) digested in aqueous/organic mobile phase with excipients at neutral pH in a 1290 LC system. And was evaluated by the integrated peak. Fusion proteins purified from CHO host cells are shown on SDS-PAGE depicted below.

Stability evaluation

The fusion proteins purified from the two formulations were partially-aliquoted into sterile tubes and stressed by 5 freeze-thaw cycles including more than 1 hour at -80°C and room temperature, respectively, or by incubation at 37°C for 1 week. I did. Stressed samples were evaluated for concentration and turbidity by UV spectroscopy using a UV transparent 96 well plate (Corning 3635) with SpectraMax M2 and SoftMaxPro software (Molecular Devices), SDS-PAGE and analytical SEC, and control non-stressed Compared to the same analysis of the sample. Analysis of control and stress samples for a single exemplary ProTriTAC molecule purified from 293 host cells Overlay of chromatograms from SEC is depicted below.

The result was that ProTriTAC was produced in a yield comparable to normal TriTAC from a CHO stable pool; And the protein was stable after repeated freeze-thaw for 1 week and 37°C.

D. Demonstration of functional masking and stability of ProTriTAC in a 3-week cynomolgus monkey pharmacokinetic study in vivo

A single dose of PSMA-targeted ProTriTAC (SEQ ID NO: 119), non-cleavable ProTriTAC (SEQ ID NO: 120), unmasked/non-cleavable TriTAC (SEQ ID NO: 123) and protease-activated ProTriTAC (SEQ ID NO: 121). The mimicked active drug was administered to cynomolgus monkeys at 0.1 mg/kg via intravenous injection. Plasma samples were collected at labeled time points. ProTriTAC concentrations were determined using a ligand binding assay with biotinylated recombinant human PSMA (R&D system) and 11D3 cloned sulfo-tagged anti-CD3 idiotype antibody in MSD analysis (Meso Scale Diagnostic, LLC). Pharmacokinetic parameters were estimated using Phoenix WinNonlin pharmacokinetic software using a non-compartmental approach consistent with the intravenous bolus route of administration.

To calculate the prodrug conversion in vivo, P is the concentration of the prodrug, A is the concentration of the active drug, k _a is the rate of prodrug activation in circulation, k _c,P is the clearance rate of the prodrug and k _{c ,A} is the clearance rate of the active drug, and the system of the following differential equation was solved to estimate the concentration of the active drug in circulation.

The clearance of prodrug, active drug and non-cleavable prodrug control (k _{c, NCLV} ) was determined empirically in cynomolgus monkeys. To estimate the rate of prodrug activation in circulation, the inventors have assumed that the difference between the clearance rates of cleavable prodrugs and non-cleavable prodrugs occurs only with non-specific activation in circulation. Therefore, the conversion rate of prodrug into active drug in circulation was estimated by subtracting the clearance rate of cleavable prodrug from non-cleavable prodrug.

The initial concentration of prodrug in circulation was determined empirically and the initial concentration of active drug was assumed to be zero.

Results and discussion

The results of Example 13 show that the therapeutic activity is masked by a masking domain that binds both serum albumin and active polypeptide, a cytokine or a functional fragment or mutein thereof or a poly with the desired therapeutic activity such as anti-CD3 scFV. It indicates that a fusion protein containing a peptide can be prepared. The masking domain is operably linked to the active domain through a protease cleavable linker. The results indicate that this type of fusion protein can be administered as an inactive protein that is activated upon protease cleavage at the desired site of therapeutic activity, such as in tumors.

The amino acid sequence of the fusion protein used in Example 13 is provided as SEQ ID NO: 116-123.

Sample fusion protein constructs are detailed in Table 3. In Table 3, "L" is an abbreviation of "linker" and "cleav. link." is an abbreviation of "cleavable linker". The other abbreviation “mIFNg” stands for mouse interferon gamma (IFNg); “hAlbumin” stands for human serum albumin (HSA); “mAlbumin” refers to mouse serum albumin.

Integration by reference

The entire disclosures of all patent and non-patent publications cited herein are incorporated by reference in their entirety, respectively, for all purposes.

Another embodiment

The disclosure presented above may cover a number of distinct inventions of independent utility. While each of these inventions has been disclosed in a preferred form(s), the specific embodiments disclosed and illustrated herein are not to be considered in a limiting sense because numerous modifications are possible. The subject matter of the present invention includes all novel and obscure combinations and subcombinations of the various elements, features, functions and/or characteristics disclosed herein. The following claims specifically point out certain combinations and subcombinations which are considered novel and obscure. Invention embodied in other combinations and subcombinations of features, functions, elements and/or features may be claimed in this application, in an application claiming priority in this application, or in a related application. These claims, whether related to different inventions or to the same invention, and whether broader, narrower, identical, or different in scope compared to the original claims, also cover the scope of the invention of the present disclosure. It is considered included within the subject.

SEQUENCE LISTING <110> WEREWOLF THERAPEUTICS, INC. <120> ACTIVATABLE INTERLEUKIN-2 POLYPEPTIDES AND METHODS OF USE THEREOF <130> 105365-0021 <140> <141> <150> 62/756,507 <151> 2018-11-06 <150> 62/756,504 <151> 2018-11-06 <150> 62/671,225 <151> 2018-05-14 <160> 137 <170> PatentIn version 3.5 <210> 1 <211> 153 <212> PRT <213> Homo sapiens <400> 1 Met Tyr Arg Met Gln Leu Leu Ser Cys Ile Ala Leu Ser Leu Ala Leu 1 5 10 15 Val Thr Asn Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu 20 25 30 Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile 35 40 45 Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe 50 55 60 Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu 65 70 75 80 Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys 85 90 95 Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile 100 105 110 Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala 115 120 125 Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe 130 135 140 Cys Gln Ser Ile Ile Ser Thr Leu Thr 145 150 <210> 2 <211> 609 <212> PRT <213> Homo sapiens <400> 2 Met Lys Trp Val Thr Phe Ile Ser Leu Leu Phe Leu Phe Ser Ser Ala 1 5 10 15 Tyr Ser Arg Gly Val Phe Arg Arg Asp Ala His Lys Ser Glu Val Ala 20 25 30 His Arg Phe Lys Asp Leu Gly Glu Glu Asn Phe Lys Ala Leu Val Leu 35 40 45 Ile Ala Phe Ala Gln Tyr Leu Gln Gln Cys Pro Phe Glu Asp His Val 50 55 60 Lys Leu Val Asn Glu Val Thr Glu Phe Ala Lys Thr Cys Val Ala Asp 65 70 75 80 Glu Ser Ala Glu Asn Cys Asp Lys Ser Leu His Thr Leu Phe Gly Asp 85 90 95 Lys Leu Cys Thr Val Ala Thr Leu Arg Glu Thr Tyr Gly Glu Met Ala 100 105 110 Asp Cys Cys Ala Lys Gln Glu Pro Glu Arg Asn Glu Cys Phe Leu Gln 115 120 125 His Lys Asp Asp Asn Pro Asn Leu Pro Arg Leu Val Arg Pro Glu Val 130 135 140 Asp Val Met Cys Thr Ala Phe His Asp Asn Glu Glu Thr Phe Leu Lys 145 150 155 160 Lys Tyr Leu Tyr Glu Ile Ala Arg Arg His Pro Tyr Phe Tyr Ala Pro 165 170 175 Glu Leu Leu Phe Phe Ala Lys Arg Tyr Lys Ala Ala Phe Thr Glu Cys 180 185 190 Cys Gln Ala Ala Asp Lys Ala Ala Cys Leu Leu Pro Lys Leu Asp Glu 195 200 205 Leu Arg Asp Glu Gly Lys Ala Ser Ser Ala Lys Gln Gly Leu Lys Cys 210 215 220 Ala Ser Leu Gln Lys Phe Gly Glu Arg Ala Phe Lys Ala Trp Ala Val 225 230 235 240 Ala Arg Leu Ser Gln Arg Phe Pro Lys Ala Glu Phe Ala Glu Val Ser 245 250 255 Lys Leu Val Thr Asp Leu Thr Lys Val His Thr Glu Cys Cys His Gly 260 265 270 Asp Leu Leu Glu Cys Ala Asp Asp Arg Ala Asp Leu Ala Lys Tyr Ile 275 280 285 Cys Glu Asn Gln Asp Ser Ile Ser Ser Lys Leu Lys Glu Cys Cys Glu 290 295 300 Lys Pro Leu Leu Glu Lys Ser His Cys Ile Ala Glu Val Glu Asn Asp 305 310 315 320 Glu Met Pro Ala Asp Leu Pro Ser Leu Ala Ala Asp Phe Val Gly Ser 325 330 335 Lys Asp Val Cys Lys Asn Tyr Ala Glu Ala Lys Asp Val Phe Leu Gly 340 345 350 Met Phe Leu Tyr Glu Tyr Ala Arg Arg His Pro Asp Tyr Ser Val Val 355 360 365 Leu Leu Leu Arg Leu Ala Lys Thr Tyr Glu Thr Thr Leu Glu Lys Cys 370 375 380 Cys Ala Ala Ala Asp Pro His Glu Cys Tyr Ala Lys Val Phe Asp Glu 385 390 395 400 Phe Lys Pro Leu Val Glu Glu Pro Gln Asn Leu Ile Lys Gln Asn Cys 405 410 415 Glu Leu Phe Glu Gln Leu Gly Glu Tyr Lys Phe Gln Asn Ala Leu Leu 420 425 430 Val Arg Tyr Thr Lys Lys Val Pro Gln Val Ser Thr Pro Thr Leu Val 435 440 445 Glu Val Ser Arg Asn Leu Gly Lys Val Gly Ser Lys Cys Cys Lys His 450 455 460 Pro Glu Ala Lys Arg Met Pro Cys Ala Glu Asp Cys Leu Ser Val Phe 465 470 475 480 Leu Asn Gln Leu Cys Val Leu His Glu Lys Thr Pro Val Ser Asp Arg 485 490 495 Val Thr Lys Cys Cys Thr Glu Ser Leu Val Asn Gly Arg Pro Cys Phe 500 505 510 Ser Ala Leu Glu Val Asp Glu Thr Tyr Val Pro Lys Glu Phe Asn Ala 515 520 525 Glu Thr Phe Thr Phe His Ala Asp Ile Cys Thr Leu Ser Glu Lys Glu 530 535 540 Arg Gln Ile Lys Lys Gln Thr Ala Leu Val Glu Leu Val Lys His Lys 545 550 555 560 Pro Lys Ala Thr Lys Glu Gln Leu Lys Ala Val Met Asp Asp Phe Ala 565 570 575 Ala Phe Val Glu Lys Cys Cys Lys Ala Asp Asp Lys Glu Thr Cys Phe 580 585 590 Ala Glu Glu Gly Lys Lys Leu Val Ala Ala Ser Gln Ala Ala Leu Gly 595 600 605 Leu <210> 3 <211> 8 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: MMP7 cleavage domain sequence" <400> 3 Lys Arg Ala Leu Gly Leu Pro Gly 1 5 <210> 4 <211> 40 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: MMP7 cleavage domain sequence" <400> 4 Asp Glu Asp Glu Asp Glu Asp Glu Asp Glu Asp Glu Asp Glu Asp Glu 1 5 10 15 Arg Pro Leu Ala Leu Trp Arg Ser Asp Arg Asp Arg Asp Arg Asp Arg 20 25 30 Asp Arg Asp Arg Asp Arg Asp Arg 35 40 <210> 5 <211> 5 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: MMP9 cleavage domain sequence" <220> <221> VARIANT <222> (3)..(3) <223> /replace="Thr" <220> <221> VARIANT <222> (4)..(4) <223> /replace="Ile" <220> <221> VARIANT <222> (5)..(5) <223> /replace="Thr" <220> <221> SITE <222> (1)..(5) <223> /note="Variant residues given in the sequence have no preference with respect to those in the annotations for variant positions" <400> 5 Pro Arg Ser Leu Ser 1 5 <210> 6 <211> 5 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: MMP9 cleavage domain sequence" <400> 6 Leu Glu Ala Thr Ala 1 5 <210> 7 <211> 10 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: MMP11 cleavage domain sequence" <400> 7 Gly Gly Ala Ala Asn Leu Val Arg Gly Gly 1 5 10 <210> 8 <211> 10 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: MMP14 cleavage domain sequence" <400> 8 Ser Gly Arg Ile Gly Phe Leu Arg Thr Ala 1 5 10 <210> 9 <211> 6 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: MMP cleavage domain sequence" <400> 9 Pro Leu Gly Leu Ala Gly 1 5 <210> 10 <211> 6 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: MMP cleavage domain sequence" <220> <221> MOD_RES <222> (6)..(6) <223> Any amino acid <400> 10 Pro Leu Gly Leu Ala Xaa 1 5 <210> 11 <211> 6 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: MMP cleavage domain sequence" <220> <221> MOD_RES <222> (4)..(4) <223> Cys(me) <400> 11 Pro Leu Gly Xaa Ala Gly 1 5 <210> 12 <211> 8 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: MMP cleavage domain sequence" <400> 12 Glu Ser Pro Ala Tyr Tyr Thr Ala 1 5 <210> 13 <211> 6 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: MMP cleavage domain sequence" <400> 13 Arg Leu Gln Leu Lys Leu 1 5 <210> 14 <211> 7 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: MMP cleavage domain sequence" <400> 14 Arg Leu Gln Leu Lys Ala Cys 1 5 <210> 15 <211> 7 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: MMP2, MMP9, MMP14 cleavage domain sequence" <220> <221> MOD_RES <222> (3)..(3) <223> Cit <220> <221> MOD_RES <222> (5)..(5) <223> Hof <400> 15 Glu Pro Xaa Gly Xaa Tyr Leu 1 5 <210> 16 <211> 5 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: Urokinase plasminogen activator (uPA) cleavage domain sequence" <400> 16 Ser Gly Arg Ser Ala 1 5 <210> 17 <211> 4 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: Urokinase plasminogen activator (uPA) cleavage domain sequence" <400> 17 Asp Ala Phe Lys One <210> 18 <211> 5 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: Urokinase plasminogen activator (uPA) cleavage domain sequence" <400> 18 Gly Gly Gly Arg Arg 1 5 <210> 19 <211> 4 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: Lysosomal Enzyme cleavage domain sequence" <400> 19 Gly Phe Leu Gly One <210> 20 <211> 4 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: Lysosomal Enzyme cleavage domain sequence" <400> 20 Ala Leu Ala Leu One <210> 21 <211> 2 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: Lysosomal Enzyme cleavage domain sequence" <400> 21 Phe Lys One <210> 22 <211> 3 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: Cathepsin B cleavage domain sequence" <400> 22 Asn Leu Leu One <210> 23 <211> 5 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: Cathepsin D cleavage domain sequence" <220> <221> MOD_RES <222> (3)..(3) <223> Cys(et) <400> 23 Pro Ile Xaa Phe Phe 1 5 <210> 24 <211> 8 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: Cathepsin K cleavage domain sequence" <400> 24 Gly Gly Pro Arg Gly Leu Pro Gly 1 5 <210> 25 <211> 6 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: Prostate Specific Antigen cleavage domain sequence" <400> 25 His Ser Ser Lys Leu Gln 1 5 <210> 26 <211> 7 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: Prostate Specific Antigen cleavage domain sequence" <400> 26 His Ser Ser Lys Leu Gln Leu 1 5 <210> 27 <211> 9 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: Prostate Specific Antigen cleavage domain sequence" <400> 27 His Ser Ser Lys Leu Gln Glu Asp Ala 1 5 <210> 28 <211> 10 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: Herpes Simplex Virus Protease cleavage domain sequence" <400> 28 Leu Val Leu Ala Ser Ser Ser Phe Gly Tyr 1 5 10 <210> 29 <211> 10 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: HIV Protease cleavage domain sequence" <400> 29 Gly Val Ser Gln Asn Tyr Pro Ile Val Gly 1 5 10 <210> 30 <211> 10 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: CMV Protease cleavage domain sequence" <400> 30 Gly Val Val Gln Ala Ser Cys Arg Leu Ala 1 5 10 <210> 31 <211> 4 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: Thrombin cleavage domain sequence" <220> <221> MOD_RES <222> (2)..(2) <223> 2-carboxy piperdine <400> 31 Phe Xaa Arg Ser One <210> 32 <211> 6 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: Thrombin cleavage domain sequence" <400> 32 Asp Pro Arg Ser Phe Leu 1 5 <210> 33 <211> 6 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: Thrombin cleavage domain sequence" <400> 33 Pro Pro Arg Ser Phe Leu 1 5 <210> 34 <211> 4 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: Caspase-3 cleavage domain sequence" <400> 34 Asp Glu Val Asp One <210> 35 <211> 5 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: Caspase-3 cleavage domain sequence" <400> 35 Asp Glu Val Asp Pro 1 5 <210> 36 <211> 8 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: Caspase-3 cleavage domain sequence" <400> 36 Lys Gly Ser Gly Asp Val Glu Gly 1 5 <210> 37 <211> 6 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: Interleukin 1-Beta converting enzyme cleavage domain sequence" <400> 37 Gly Trp Glu His Asp Gly 1 5 <210> 38 <211> 7 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: Enterokinase cleavage domain sequence" <400> 38 Glu Asp Asp Asp Asp Lys Ala 1 5 <210> 39 <211> 9 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: FAP cleavage domain sequence" <400> 39 Lys Gln Glu Gln Asn Pro Gly Ser Thr 1 5 <210> 40 <211> 6 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: Kallikrein 2 cleavage domain sequence" <400> 40 Gly Lys Ala Phe Arg Arg 1 5 <210> 41 <211> 4 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: Plasmin cleavage domain sequence" <400> 41 Asp Ala Phe Lys One <210> 42 <211> 4 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: Plasmin cleavage domain sequence" <400> 42 Asp Val Leu Lys One <210> 43 <211> 4 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: Plasmin cleavage domain sequence" <400> 43 Asp Ala Phe Lys One <210> 44 <211> 7 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: TOP cleavage domain sequence" <400> 44 Ala Leu Leu Leu Ala Leu Leu 1 5 <210> 45 <211> 652 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 45 Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Ile Phe Ser Ile Asp 20 25 30 Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val 35 40 45 Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser Tyr Asp Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Gly Val Tyr Tyr Cys Asn 85 90 95 Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys Gly Thr Gln Val Thr Val 100 105 110 Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 115 120 125 Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu 130 135 140 His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr 145 150 155 160 Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro 165 170 175 Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu 180 185 190 Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His 195 200 205 Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu 210 215 220 Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr 225 230 235 240 Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser 245 250 255 Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly Pro Ala Gly Met Lys 260 265 270 Gly Leu Pro Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu 275 280 285 Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe 290 295 300 Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys 305 310 315 320 Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr Leu 325 330 335 Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala 340 345 350 Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr 355 360 365 Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser Gln 370 375 380 Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly 385 390 395 400 Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly 405 410 415 Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser 420 425 430 Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg Gln Ala Pro 435 440 445 Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser Ser Tyr Thr 450 455 460 Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser Arg Asp Asn 465 470 475 480 Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp 485 490 495 Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp Ala Leu Asp 500 505 510 Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly 515 520 525 Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr 530 535 540 Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile 545 550 555 560 Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly Trp Tyr Gln 565 570 575 Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser Ala Ser Phe 580 585 590 Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr 595 600 605 Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr 610 615 620 Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe Gly Gly Gly 625 630 635 640 Thr Lys Val Glu Ile Lys His His His His His His 645 650 <210> 46 <211> 652 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 46 Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Ile Phe Ser Ile Asp 20 25 30 Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val 35 40 45 Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser Tyr Asp Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Gly Val Tyr Tyr Cys Asn 85 90 95 Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys Gly Thr Gln Val Thr Val 100 105 110 Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 115 120 125 Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly 130 135 140 Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser 145 150 155 160 Tyr Thr Leu Ala Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp 165 170 175 Val Ala Ala Ile Asp Ser Ser Ser Tyr Thr Tyr Ser Pro Asp Thr Val 180 185 190 Arg Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr 195 200 205 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 210 215 220 Ala Arg Asp Ser Asn Trp Asp Ala Leu Asp Tyr Trp Gly Gln Gly Thr 225 230 235 240 Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 245 250 255 Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu 260 265 270 Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln 275 280 285 Asn Val Gly Thr Asn Val Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala 290 295 300 Pro Lys Ala Leu Ile Tyr Ser Ala Ser Phe Arg Tyr Ser Gly Val Pro 305 310 315 320 Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile 325 330 335 Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr 340 345 350 Tyr Thr Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 355 360 365 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu 370 375 380 Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser 385 390 395 400 Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly 405 410 415 Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser 420 425 430 Ser Ile Ser Gly Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys 435 440 445 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu 450 455 460 Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr 465 470 475 480 Ile Gly Gly Ser Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val 485 490 495 Ser Ser Ser Gly Gly Pro Gly Pro Ala Gly Met Lys Gly Leu Pro Gly 500 505 510 Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu 515 520 525 His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr 530 535 540 Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro 545 550 555 560 Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu 565 570 575 Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His 580 585 590 Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu 595 600 605 Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr 610 615 620 Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser 625 630 635 640 Ile Ile Ser Thr Leu Thr His His His His His His 645 650 <210> 47 <211> 553 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 47 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Thr Leu Ala Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Ala Ile Asp Ser Ser Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Asp Ser Asn Trp Asp Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr 100 105 110 Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 115 120 125 Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser 130 135 140 Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn 145 150 155 160 Val Gly Thr Asn Val Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro 165 170 175 Lys Ala Leu Ile Tyr Ser Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser 180 185 190 Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser 195 200 205 Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Tyr Cys Gln Gln Tyr Tyr 210 215 220 Thr Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Gly 225 230 235 240 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 245 250 255 Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Gly Gly 260 265 270 Pro Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Ala Pro Thr Ser 275 280 285 Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp 290 295 300 Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu 305 310 315 320 Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu 325 330 335 Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu 340 345 350 Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp 355 360 365 Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu 370 375 380 Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu 385 390 395 400 Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu 405 410 415 Thr Ser Gly Gly Pro Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser 420 425 430 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn 435 440 445 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe 450 455 460 Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 465 470 475 480 Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val 485 490 495 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr 500 505 510 Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 515 520 525 Thr Ile Gly Gly Ser Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr 530 535 540 Val Ser Ser His His His His His His 545 550 <210> 48 <211> 553 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 48 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Thr Leu Ala Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Ala Ile Asp Ser Ser Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Asp Ser Asn Trp Asp Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr 100 105 110 Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 115 120 125 Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser 130 135 140 Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn 145 150 155 160 Val Gly Thr Asn Val Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro 165 170 175 Lys Ala Leu Ile Tyr Ser Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser 180 185 190 Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser 195 200 205 Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Tyr Cys Gln Gln Tyr Tyr 210 215 220 Thr Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Gly 225 230 235 240 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 245 250 255 Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln 260 265 270 Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg 275 280 285 Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser 290 295 300 Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile 305 310 315 320 Ser Gly Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg 325 330 335 Phe Thr Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met 340 345 350 Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly 355 360 365 Gly Ser Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser 370 375 380 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser 385 390 395 400 Gly Gly Pro Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Ala Pro 405 410 415 Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu 420 425 430 Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro 435 440 445 Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala 450 455 460 Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu 465 470 475 480 Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro 485 490 495 Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly 500 505 510 Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile 515 520 525 Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile Ile Ser 530 535 540 Thr Leu Thr His His His His His His 545 550 <210> 49 <211> 553 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 49 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly Pro Ala Gly Met Lys Gly 130 135 140 Leu Pro Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val 145 150 155 160 Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr 165 170 175 Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly 180 185 190 Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr Leu Tyr 195 200 205 Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys 210 215 220 Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala 225 230 235 240 Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser Gln Gly 245 250 255 Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 260 265 270 Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 275 280 285 Gly Gly Gly Gly Ser Ser Gly Gly Pro Gly Pro Ala Gly Met Lys Gly 290 295 300 Leu Pro Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val 305 310 315 320 Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr 325 330 335 Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg Gln Ala Pro Gly Lys Gly 340 345 350 Leu Glu Trp Val Ala Ala Ile Asp Ser Ser Ser Tyr Thr Tyr Ser Pro 355 360 365 Asp Thr Val Arg Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn 370 375 380 Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val 385 390 395 400 Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp Ala Leu Asp Tyr Trp Gly 405 410 415 Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly 420 425 430 Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro 435 440 445 Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Lys 450 455 460 Ala Ser Gln Asn Val Gly Thr Asn Val Gly Trp Tyr Gln Gln Lys Pro 465 470 475 480 Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser Ala Ser Phe Arg Tyr Ser 485 490 495 Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr 500 505 510 Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys 515 520 525 Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Val 530 535 540 Glu Ile Lys His His His His His His 545 550 <210> 50 <211> 682 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 50 Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Ile Phe Ser Ile Asp 20 25 30 Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val 35 40 45 Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser Tyr Asp Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Gly Val Tyr Tyr Cys Asn 85 90 95 Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys Gly Thr Gln Val Thr Val 100 105 110 Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 115 120 125 Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu 130 135 140 His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr 145 150 155 160 Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro 165 170 175 Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu 180 185 190 Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His 195 200 205 Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu 210 215 220 Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr 225 230 235 240 Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser 245 250 255 Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly Pro Ala Gly Met Lys 260 265 270 Gly Leu Pro Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu 275 280 285 Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe 290 295 300 Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys 305 310 315 320 Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr Leu 325 330 335 Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala 340 345 350 Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr 355 360 365 Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser Gln 370 375 380 Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly 385 390 395 400 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 405 410 415 Ser Gly Gly Gly Gly Ser Ser Gly Gly Pro Gly Pro Ala Gly Met Lys 420 425 430 Gly Leu Pro Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu 435 440 445 Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe 450 455 460 Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg Gln Ala Pro Gly Lys 465 470 475 480 Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser Ser Tyr Thr Tyr Ser 485 490 495 Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys 500 505 510 Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala 515 520 525 Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp Ala Leu Asp Tyr Trp 530 535 540 Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly 545 550 555 560 Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser 565 570 575 Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys 580 585 590 Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly Trp Tyr Gln Gln Lys 595 600 605 Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser Ala Ser Phe Arg Tyr 610 615 620 Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe 625 630 635 640 Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr 645 650 655 Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys 660 665 670 Val Glu Ile Lys His His His His His His 675 680 <210> 51 <211> 667 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 51 Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Ile Phe Ser Ile Asp 20 25 30 Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val 35 40 45 Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser Tyr Asp Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Gly Val Tyr Tyr Cys Asn 85 90 95 Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys Gly Thr Gln Val Thr Val 100 105 110 Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 115 120 125 Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu 130 135 140 His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr 145 150 155 160 Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro 165 170 175 Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu 180 185 190 Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His 195 200 205 Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu 210 215 220 Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr 225 230 235 240 Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser 245 250 255 Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly Pro Ala Gly Met Lys 260 265 270 Gly Leu Pro Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu 275 280 285 Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe 290 295 300 Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys 305 310 315 320 Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr Leu 325 330 335 Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala 340 345 350 Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr 355 360 365 Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser Gln 370 375 380 Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly 385 390 395 400 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 405 410 415 Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly 420 425 430 Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly 435 440 445 Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg Gln Ala Pro Gly 450 455 460 Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser Ser Tyr Thr Tyr 465 470 475 480 Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala 485 490 495 Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr 500 505 510 Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp Ala Leu Asp Tyr 515 520 525 Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ser Gly Gly Pro Gly 530 535 540 Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Asp Ile Gln Met Thr Gln 545 550 555 560 Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr 565 570 575 Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly Trp Tyr Gln Gln 580 585 590 Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser Ala Ser Phe Arg 595 600 605 Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp 610 615 620 Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr 625 630 635 640 Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe Gly Gly Gly Thr 645 650 655 Lys Val Glu Ile Lys His His His His His His 660 665 <210> 52 <211> 682 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 52 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly Pro Ala Gly Met Lys Gly 130 135 140 Leu Pro Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 145 150 155 160 Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 165 170 175 Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro 180 185 190 Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser 195 200 205 Ser Tyr Thr Leu Ala Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu 210 215 220 Trp Val Ala Ala Ile Asp Ser Ser Ser Tyr Thr Tyr Ser Pro Asp Thr 225 230 235 240 Val Arg Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu 245 250 255 Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr 260 265 270 Cys Ala Arg Asp Ser Asn Trp Asp Ala Leu Asp Tyr Trp Gly Gln Gly 275 280 285 Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 290 295 300 Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser 305 310 315 320 Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Lys Ala Ser 325 330 335 Gln Asn Val Gly Thr Asn Val Gly Trp Tyr Gln Gln Lys Pro Gly Lys 340 345 350 Ala Pro Lys Ala Leu Ile Tyr Ser Ala Ser Phe Arg Tyr Ser Gly Val 355 360 365 Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 370 375 380 Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln 385 390 395 400 Tyr Tyr Thr Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Val Glu Ile 405 410 415 Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 420 425 430 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn 435 440 445 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe 450 455 460 Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 465 470 475 480 Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val 485 490 495 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr 500 505 510 Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 515 520 525 Thr Ile Gly Gly Ser Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr 530 535 540 Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 545 550 555 560 Gly Ser Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Leu Val Gln Ala 565 570 575 Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Ile Phe Ser 580 585 590 Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu 595 600 605 Leu Val Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser Tyr Asp Asp Ser 610 615 620 Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val 625 630 635 640 Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Gly Val Tyr Tyr 645 650 655 Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys Gly Thr Gln Val 660 665 670 Thr Val Ser Ser His His His His His His 675 680 <210> 53 <211> 393 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 53 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly Pro Ala Gly Met Lys Gly 130 135 140 Leu Pro Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val 145 150 155 160 Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr 165 170 175 Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg Gln Ala Pro Gly Lys Gly 180 185 190 Leu Glu Trp Val Ala Ala Ile Asp Ser Ser Ser Tyr Thr Tyr Ser Pro 195 200 205 Asp Thr Val Arg Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn 210 215 220 Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val 225 230 235 240 Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp Ala Leu Asp Tyr Trp Gly 245 250 255 Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly 260 265 270 Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro 275 280 285 Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Lys 290 295 300 Ala Ser Gln Asn Val Gly Thr Asn Val Gly Trp Tyr Gln Gln Lys Pro 305 310 315 320 Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser Ala Ser Phe Arg Tyr Ser 325 330 335 Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr 340 345 350 Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys 355 360 365 Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Val 370 375 380 Glu Ile Lys His His His His His His 385 390 <210> 54 <211> 423 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 54 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly Pro Ala Gly Met Lys Gly 130 135 140 Leu Pro Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 145 150 155 160 Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 165 170 175 Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro 180 185 190 Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser 195 200 205 Ser Tyr Thr Leu Ala Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu 210 215 220 Trp Val Ala Ala Ile Asp Ser Ser Ser Tyr Thr Tyr Ser Pro Asp Thr 225 230 235 240 Val Arg Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu 245 250 255 Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr 260 265 270 Cys Ala Arg Asp Ser Asn Trp Asp Ala Leu Asp Tyr Trp Gly Gln Gly 275 280 285 Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 290 295 300 Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser 305 310 315 320 Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Lys Ala Ser 325 330 335 Gln Asn Val Gly Thr Asn Val Gly Trp Tyr Gln Gln Lys Pro Gly Lys 340 345 350 Ala Pro Lys Ala Leu Ile Tyr Ser Ala Ser Phe Arg Tyr Ser Gly Val 355 360 365 Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 370 375 380 Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln 385 390 395 400 Tyr Tyr Thr Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Val Glu Ile 405 410 415 Lys His His His His His His 420 <210> 55 <211> 682 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 55 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly Pro Ala Gly Met Lys Gly 130 135 140 Leu Pro Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 145 150 155 160 Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 165 170 175 Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro 180 185 190 Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser 195 200 205 Ser Tyr Thr Leu Ala Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu 210 215 220 Trp Val Ala Ala Ile Asp Ser Ser Ser Tyr Thr Tyr Ser Pro Asp Thr 225 230 235 240 Val Arg Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu 245 250 255 Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr 260 265 270 Cys Ala Arg Asp Ser Asn Trp Asp Ala Leu Asp Tyr Trp Gly Gln Gly 275 280 285 Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 290 295 300 Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser 305 310 315 320 Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Lys Ala Ser 325 330 335 Gln Asn Val Gly Thr Asn Val Gly Trp Tyr Gln Gln Lys Pro Gly Lys 340 345 350 Ala Pro Lys Ala Leu Ile Tyr Ser Ala Ser Phe Arg Tyr Ser Gly Val 355 360 365 Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 370 375 380 Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln 385 390 395 400 Tyr Tyr Thr Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Val Glu Ile 405 410 415 Lys Ser Gly Gly Pro Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser 420 425 430 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn 435 440 445 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe 450 455 460 Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 465 470 475 480 Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val 485 490 495 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr 500 505 510 Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 515 520 525 Thr Ile Gly Gly Ser Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr 530 535 540 Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 545 550 555 560 Gly Ser Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Leu Val Gln Ala 565 570 575 Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Ile Phe Ser 580 585 590 Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu 595 600 605 Leu Val Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser Tyr Asp Asp Ser 610 615 620 Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val 625 630 635 640 Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Gly Val Tyr Tyr 645 650 655 Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys Gly Thr Gln Val 660 665 670 Thr Val Ser Ser His His His His His His 675 680 <210> 56 <211> 682 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 56 Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Leu Ala Gln Ala Gly Gly 1 5 10 15 Ser Leu Ser Leu Ser Cys Ala Ala Ser Gly Phe Thr Val Ser Asn Ser 20 25 30 Val Met Ala Trp Tyr Arg Gln Thr Pro Gly Lys Gln Arg Glu Phe Val 35 40 45 Ala Ile Ile Asn Ser Val Gly Ser Thr Asn Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu 65 70 75 80 Gln Met Asn Asn Leu Lys Pro Glu Asp Thr Ala Val Tyr Val Cys Asn 85 90 95 Arg Asn Phe Asp Arg Ile Tyr Trp Gly Gln Gly Thr Gln Val Thr Val 100 105 110 Ser Ser Ser Gly Gly Pro Gly Pro Ala Gly Met Lys Gly Leu Pro Gly 115 120 125 Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly 130 135 140 Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser 145 150 155 160 Tyr Thr Leu Ala Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp 165 170 175 Val Ala Ala Ile Asp Ser Ser Ser Tyr Thr Tyr Ser Pro Asp Thr Val 180 185 190 Arg Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr 195 200 205 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 210 215 220 Ala Arg Asp Ser Asn Trp Asp Ala Leu Asp Tyr Trp Gly Gln Gly Thr 225 230 235 240 Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 245 250 255 Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu 260 265 270 Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln 275 280 285 Asn Val Gly Thr Asn Val Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala 290 295 300 Pro Lys Ala Leu Ile Tyr Ser Ala Ser Phe Arg Tyr Ser Gly Val Pro 305 310 315 320 Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile 325 330 335 Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr 340 345 350 Tyr Thr Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 355 360 365 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 370 375 380 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Gly 385 390 395 400 Gly Pro Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln 405 410 415 Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg 420 425 430 Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser 435 440 445 Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile 450 455 460 Ser Gly Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg 465 470 475 480 Phe Thr Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met 485 490 495 Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly 500 505 510 Gly Ser Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser 515 520 525 Ser Gly Gly Pro Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Ala 530 535 540 Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu 545 550 555 560 Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn 565 570 575 Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys 580 585 590 Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro 595 600 605 Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg 610 615 620 Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys 625 630 635 640 Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr 645 650 655 Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile Ile 660 665 670 Ser Thr Leu Thr His His His His His His 675 680 <210> 57 <211> 393 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 57 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Thr Leu Ala Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Ala Ile Asp Ser Ser Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Asp Ser Asn Trp Asp Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr 100 105 110 Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 115 120 125 Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser 130 135 140 Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn 145 150 155 160 Val Gly Thr Asn Val Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro 165 170 175 Lys Ala Leu Ile Tyr Ser Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser 180 185 190 Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser 195 200 205 Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Tyr Cys Gln Gln Tyr Tyr 210 215 220 Thr Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Ser 225 230 235 240 Gly Gly Pro Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Ala Pro 245 250 255 Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu 260 265 270 Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro 275 280 285 Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala 290 295 300 Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu 305 310 315 320 Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro 325 330 335 Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly 340 345 350 Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile 355 360 365 Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile Ile Ser 370 375 380 Thr Leu Thr His His His His His His 385 390 <210> 58 <211> 423 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 58 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Thr Leu Ala Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Ala Ile Asp Ser Ser Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Asp Ser Asn Trp Asp Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr 100 105 110 Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 115 120 125 Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser 130 135 140 Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn 145 150 155 160 Val Gly Thr Asn Val Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro 165 170 175 Lys Ala Leu Ile Tyr Ser Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser 180 185 190 Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser 195 200 205 Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Tyr Cys Gln Gln Tyr Tyr 210 215 220 Thr Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Gly 225 230 235 240 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 245 250 255 Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Gly Gly 260 265 270 Pro Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Ala Pro Thr Ser 275 280 285 Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp 290 295 300 Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu 305 310 315 320 Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu 325 330 335 Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu 340 345 350 Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp 355 360 365 Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu 370 375 380 Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu 385 390 395 400 Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu 405 410 415 Thr His His His His His His 420 <210> 59 <211> 669 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 59 Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Ile Phe Ser Ile Asp 20 25 30 Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val 35 40 45 Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser Tyr Asp Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Gly Val Tyr Tyr Cys Asn 85 90 95 Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys Gly Thr Gln Val Thr Val 100 105 110 Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 115 120 125 Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu 130 135 140 His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr 145 150 155 160 Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro 165 170 175 Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu 180 185 190 Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His 195 200 205 Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu 210 215 220 Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr 225 230 235 240 Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser 245 250 255 Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly Pro Ala Gly Met Lys 260 265 270 Gly Leu Pro Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu 275 280 285 Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe 290 295 300 Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys 305 310 315 320 Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr Leu 325 330 335 Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala 340 345 350 Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr 355 360 365 Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser Gln 370 375 380 Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly 385 390 395 400 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu 405 410 415 Gln Gln Ser Gly Ala Glu Leu Val Arg Pro Gly Thr Ser Val Lys Val 420 425 430 Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr Leu Ile Glu Trp 435 440 445 Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile Gly Val Ile Asn 450 455 460 Pro Gly Ser Gly Gly Thr Asn Tyr Asn Glu Lys Phe Lys Gly Lys Ala 465 470 475 480 Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr Met Gln Leu Ser 485 490 495 Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe Cys Ala Arg Trp Arg 500 505 510 Gly Asp Gly Tyr Tyr Ala Tyr Phe Asp Val Trp Gly Ala Gly Thr Thr 515 520 525 Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 530 535 540 Gly Gly Gly Ser Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala 545 550 555 560 Val Ser Leu Gly Gln Arg Ala Thr Ile Ser Cys Lys Ala Ser Gln Ser 565 570 575 Val Asp Tyr Asp Gly Asp Ser Tyr Met Asn Trp Tyr Gln Gln Lys Pro 580 585 590 Gly Gln Pro Pro Lys Leu Leu Ile Tyr Ala Ala Ser Asn Leu Glu Ser 595 600 605 Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr 610 615 620 Leu Asn Ile His Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr Tyr Cys 625 630 635 640 Gln Gln Ser Asn Glu Asp Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu 645 650 655 Glu Ile Lys His His His His His His Glu Pro Glu Ala 660 665 <210> 60 <211> 669 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 60 Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Ile Phe Ser Ile Asp 20 25 30 Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val 35 40 45 Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser Tyr Asp Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Gly Val Tyr Tyr Cys Asn 85 90 95 Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys Gly Thr Gln Val Thr Val 100 105 110 Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 115 120 125 Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu 130 135 140 His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr 145 150 155 160 Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro 165 170 175 Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu 180 185 190 Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His 195 200 205 Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu 210 215 220 Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr 225 230 235 240 Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser 245 250 255 Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly Pro Ala Gly Met Lys 260 265 270 Gly Leu Pro Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu 275 280 285 Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe 290 295 300 Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys 305 310 315 320 Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr Leu 325 330 335 Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala 340 345 350 Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr 355 360 365 Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser Gln 370 375 380 Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly 385 390 395 400 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Val Leu 405 410 415 Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly Gln Arg Ala Thr 420 425 430 Ile Ser Cys Lys Ala Ser Gln Ser Val Asp Tyr Asp Gly Asp Ser Tyr 435 440 445 Met Asn Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile 450 455 460 Tyr Ala Ala Ser Asn Leu Glu Ser Gly Ile Pro Ala Arg Phe Ser Gly 465 470 475 480 Ser Gly Ser Gly Thr Asp Phe Thr Leu Asn Ile His Pro Val Glu Glu 485 490 495 Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Ser Asn Glu Asp Pro Tyr 500 505 510 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Gly Gly Gly Gly Ser 515 520 525 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Gln 530 535 540 Ser Gly Ala Glu Leu Val Arg Pro Gly Thr Ser Val Lys Val Ser Cys 545 550 555 560 Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr Leu Ile Glu Trp Val Lys 565 570 575 Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile Gly Val Ile Asn Pro Gly 580 585 590 Ser Gly Gly Thr Asn Tyr Asn Glu Lys Phe Lys Gly Lys Ala Thr Leu 595 600 605 Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu 610 615 620 Thr Ser Asp Asp Ser Ala Val Tyr Phe Cys Ala Arg Trp Arg Gly Asp 625 630 635 640 Gly Tyr Tyr Ala Tyr Phe Asp Val Trp Gly Ala Gly Thr Thr Val Thr 645 650 655 Val Ser Ser His His His His His His Glu Pro Glu Ala 660 665 <210> 61 <211> 689 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 61 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly Pro Ala Gly Met Lys Gly 130 135 140 Leu Pro Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 145 150 155 160 Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln 165 170 175 Leu Gln Gln Ser Gly Ala Glu Leu Val Arg Pro Gly Thr Ser Val Lys 180 185 190 Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr Leu Ile Glu 195 200 205 Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile Gly Val Ile 210 215 220 Asn Pro Gly Ser Gly Gly Thr Asn Tyr Asn Glu Lys Phe Lys Gly Lys 225 230 235 240 Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr Met Gln Leu 245 250 255 Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe Cys Ala Arg Trp 260 265 270 Arg Gly Asp Gly Tyr Tyr Ala Tyr Phe Asp Val Trp Gly Ala Gly Thr 275 280 285 Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 290 295 300 Gly Gly Gly Gly Ser Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu 305 310 315 320 Ala Val Ser Leu Gly Gln Arg Ala Thr Ile Ser Cys Lys Ala Ser Gln 325 330 335 Ser Val Asp Tyr Asp Gly Asp Ser Tyr Met Asn Trp Tyr Gln Gln Lys 340 345 350 Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Ala Ala Ser Asn Leu Glu 355 360 365 Ser Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe 370 375 380 Thr Leu Asn Ile His Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr Tyr 385 390 395 400 Cys Gln Gln Ser Asn Glu Asp Pro Tyr Thr Phe Gly Gly Gly Thr Lys 405 410 415 Leu Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 420 425 430 Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln 435 440 445 Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe 450 455 460 Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu 465 470 475 480 Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr Leu Tyr Ala 485 490 495 Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Thr 500 505 510 Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val 515 520 525 Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser Gln Gly Thr 530 535 540 Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 545 550 555 560 Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Leu 565 570 575 Val Gln Ala Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg 580 585 590 Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly Lys 595 600 605 Gln Arg Glu Leu Val Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser Tyr 610 615 620 Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys 625 630 635 640 Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Gly 645 650 655 Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys Gly 660 665 670 Thr Gln Val Thr Val Ser Ser His His His His His His Glu Pro Glu 675 680 685 Ala <210> 62 <211> 689 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 62 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly Pro Ala Gly Met Lys Gly 130 135 140 Leu Pro Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 145 150 155 160 Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Val 165 170 175 Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly Gln Arg Ala 180 185 190 Thr Ile Ser Cys Lys Ala Ser Gln Ser Val Asp Tyr Asp Gly Asp Ser 195 200 205 Tyr Met Asn Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu 210 215 220 Ile Tyr Ala Ala Ser Asn Leu Glu Ser Gly Ile Pro Ala Arg Phe Ser 225 230 235 240 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Asn Ile His Pro Val Glu 245 250 255 Glu Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Ser Asn Glu Asp Pro 260 265 270 Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Gly Gly Gly Gly 275 280 285 Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Gln 290 295 300 Gln Ser Gly Ala Glu Leu Val Arg Pro Gly Thr Ser Val Lys Val Ser 305 310 315 320 Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr Leu Ile Glu Trp Val 325 330 335 Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile Gly Val Ile Asn Pro 340 345 350 Gly Ser Gly Gly Thr Asn Tyr Asn Glu Lys Phe Lys Gly Lys Ala Thr 355 360 365 Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr Met Gln Leu Ser Ser 370 375 380 Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe Cys Ala Arg Trp Arg Gly 385 390 395 400 Asp Gly Tyr Tyr Ala Tyr Phe Asp Val Trp Gly Ala Gly Thr Thr Val 405 410 415 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 420 425 430 Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln 435 440 445 Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe 450 455 460 Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu 465 470 475 480 Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr Leu Tyr Ala 485 490 495 Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Thr 500 505 510 Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val 515 520 525 Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser Gln Gly Thr 530 535 540 Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 545 550 555 560 Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Leu 565 570 575 Val Gln Ala Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg 580 585 590 Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly Lys 595 600 605 Gln Arg Glu Leu Val Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser Tyr 610 615 620 Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys 625 630 635 640 Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Gly 645 650 655 Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys Gly 660 665 670 Thr Gln Val Thr Val Ser Ser His His His His His His Glu Pro Glu 675 680 685 Ala <210> 63 <211> 272 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: IL2Ra sequence" <400> 63 Met Asp Ser Tyr Leu Leu Met Trp Gly Leu Leu Thr Phe Ile Met Val 1 5 10 15 Pro Gly Cys Gln Ala Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro 20 25 30 His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn 35 40 45 Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu Tyr 50 55 60 Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys 65 70 75 80 Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro 85 90 95 Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro 100 105 110 Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro 115 120 125 Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val 130 135 140 Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His 145 150 155 160 Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr His Gly Lys Thr Arg 165 170 175 Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly Glu Met Glu Thr Ser Gln 180 185 190 Phe Pro Gly Glu Glu Lys Pro Gln Ala Ser Pro Glu Gly Arg Pro Glu 195 200 205 Ser Glu Thr Ser Cys Leu Val Thr Thr Thr Asp Phe Gln Ile Gln Thr 210 215 220 Glu Met Ala Ala Thr Met Glu Thr Ser Ile Phe Thr Thr Glu Tyr Gln 225 230 235 240 Val Ala Val Ala Gly Cys Val Phe Leu Leu Ile Ser Val Leu Leu Leu 245 250 255 Ser Gly Leu Thr Trp Gln Arg Arg Gln Arg Lys Ser Arg Arg Thr Ile 260 265 270 <210> 64 <211> 551 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: IL2Rb sequence" <400> 64 Met Ala Ala Pro Ala Leu Ser Trp Arg Leu Pro Leu Leu Ile Leu Leu 1 5 10 15 Leu Pro Leu Ala Thr Ser Trp Ala Ser Ala Ala Val Asn Gly Thr Ser 20 25 30 Gln Phe Thr Cys Phe Tyr Asn Ser Arg Ala Asn Ile Ser Cys Val Trp 35 40 45 Ser Gln Asp Gly Ala Leu Gln Asp Thr Ser Cys Gln Val His Ala Trp 50 55 60 Pro Asp Arg Arg Arg Trp Asn Gln Thr Cys Glu Leu Leu Pro Val Ser 65 70 75 80 Gln Ala Ser Trp Ala Cys Asn Leu Ile Leu Gly Ala Pro Asp Ser Gln 85 90 95 Lys Leu Thr Thr Val Asp Ile Val Thr Leu Arg Val Leu Cys Arg Glu 100 105 110 Gly Val Arg Trp Arg Val Met Ala Ile Gln Asp Phe Lys Pro Phe Glu 115 120 125 Asn Leu Arg Leu Met Ala Pro Ile Ser Leu Gln Val Val His Val Glu 130 135 140 Thr His Arg Cys Asn Ile Ser Trp Glu Ile Ser Gln Ala Ser His Tyr 145 150 155 160 Phe Glu Arg His Leu Glu Phe Glu Ala Arg Thr Leu Ser Pro Gly His 165 170 175 Thr Trp Glu Glu Ala Pro Leu Leu Thr Leu Lys Gln Lys Gln Glu Trp 180 185 190 Ile Cys Leu Glu Thr Leu Thr Pro Asp Thr Gln Tyr Glu Phe Gln Val 195 200 205 Arg Val Lys Pro Leu Gln Gly Glu Phe Thr Thr Trp Ser Pro Trp Ser 210 215 220 Gln Pro Leu Ala Phe Arg Thr Lys Pro Ala Ala Leu Gly Lys Asp Thr 225 230 235 240 Ile Pro Trp Leu Gly His Leu Leu Val Gly Leu Ser Gly Ala Phe Gly 245 250 255 Phe Ile Ile Leu Val Tyr Leu Leu Ile Asn Cys Arg Asn Thr Gly Pro 260 265 270 Trp Leu Lys Lys Val Leu Lys Cys Asn Thr Pro Asp Pro Ser Lys Phe 275 280 285 Phe Ser Gln Leu Ser Ser Glu His Gly Gly Asp Val Gln Lys Trp Leu 290 295 300 Ser Ser Pro Phe Pro Ser Ser Ser Phe Ser Pro Gly Gly Leu Ala Pro 305 310 315 320 Glu Ile Ser Pro Leu Glu Val Leu Glu Arg Asp Lys Val Thr Gln Leu 325 330 335 Leu Leu Gln Gln Asp Lys Val Pro Glu Pro Ala Ser Leu Ser Ser Asn 340 345 350 His Ser Leu Thr Ser Cys Phe Thr Asn Gln Gly Tyr Phe Phe Phe His 355 360 365 Leu Pro Asp Ala Leu Glu Ile Glu Ala Cys Gln Val Tyr Phe Thr Tyr 370 375 380 Asp Pro Tyr Ser Glu Glu Asp Pro Asp Glu Gly Val Ala Gly Ala Pro 385 390 395 400 Thr Gly Ser Ser Pro Gln Pro Leu Gln Pro Leu Ser Gly Glu Asp Asp 405 410 415 Ala Tyr Cys Thr Phe Pro Ser Arg Asp Asp Leu Leu Leu Phe Ser Pro 420 425 430 Ser Leu Leu Gly Gly Pro Ser Pro Pro Ser Thr Ala Pro Gly Gly Ser 435 440 445 Gly Ala Gly Glu Glu Arg Met Pro Pro Ser Leu Gln Glu Arg Val Pro 450 455 460 Arg Asp Trp Asp Pro Gln Pro Leu Gly Pro Pro Thr Pro Gly Val Pro 465 470 475 480 Asp Leu Val Asp Phe Gln Pro Pro Pro Glu Leu Val Leu Arg Glu Ala 485 490 495 Gly Glu Glu Val Pro Asp Ala Gly Pro Arg Glu Gly Val Ser Phe Pro 500 505 510 Trp Ser Arg Pro Pro Gly Gln Gly Glu Phe Arg Ala Leu Asn Ala Arg 515 520 525 Leu Pro Leu Asn Thr Asp Ala Tyr Leu Ser Leu Gln Glu Leu Gln Gly 530 535 540 Gln Asp Pro Thr His Leu Val 545 550 <210> 65 <211> 369 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: IL2Rg sequence" <400> 65 Met Leu Lys Pro Ser Leu Pro Phe Thr Ser Leu Leu Phe Leu Gln Leu 1 5 10 15 Pro Leu Leu Gly Val Gly Leu Asn Thr Thr Ile Leu Thr Pro Asn Gly 20 25 30 Asn Glu Asp Thr Thr Ala Asp Phe Phe Leu Thr Thr Met Pro Thr Asp 35 40 45 Ser Leu Ser Val Ser Thr Leu Pro Leu Pro Glu Val Gln Cys Phe Val 50 55 60 Phe Asn Val Glu Tyr Met Asn Cys Thr Trp Asn Ser Ser Ser Glu Pro 65 70 75 80 Gln Pro Thr Asn Leu Thr Leu His Tyr Trp Tyr Lys Asn Ser Asp Asn 85 90 95 Asp Lys Val Gln Lys Cys Ser His Tyr Leu Phe Ser Glu Glu Ile Thr 100 105 110 Ser Gly Cys Gln Leu Gln Lys Lys Glu Ile His Leu Tyr Gln Thr Phe 115 120 125 Val Val Gln Leu Gln Asp Pro Arg Glu Pro Arg Arg Gln Ala Thr Gln 130 135 140 Met Leu Lys Leu Gln Asn Leu Val Ile Pro Trp Ala Pro Glu Asn Leu 145 150 155 160 Thr Leu His Lys Leu Ser Glu Ser Gln Leu Glu Leu Asn Trp Asn Asn 165 170 175 Arg Phe Leu Asn His Cys Leu Glu His Leu Val Gln Tyr Arg Thr Asp 180 185 190 Trp Asp His Ser Trp Thr Glu Gln Ser Val Asp Tyr Arg His Lys Phe 195 200 205 Ser Leu Pro Ser Val Asp Gly Gln Lys Arg Tyr Thr Phe Arg Val Arg 210 215 220 Ser Arg Phe Asn Pro Leu Cys Gly Ser Ala Gln His Trp Ser Glu Trp 225 230 235 240 Ser His Pro Ile His Trp Gly Ser Asn Thr Ser Lys Glu Asn Pro Phe 245 250 255 Leu Phe Ala Leu Glu Ala Val Val Ile Ser Val Gly Ser Met Gly Leu 260 265 270 Ile Ile Ser Leu Leu Cys Val Tyr Phe Trp Leu Glu Arg Thr Met Pro 275 280 285 Arg Ile Pro Thr Leu Lys Asn Leu Glu Asp Leu Val Thr Glu Tyr His 290 295 300 Gly Asn Phe Ser Ala Trp Ser Gly Val Ser Lys Gly Leu Ala Glu Ser 305 310 315 320 Leu Gln Pro Asp Tyr Ser Glu Arg Leu Cys Leu Val Ser Glu Ile Pro 325 330 335 Pro Lys Gly Gly Ala Leu Gly Glu Gly Pro Gly Ala Ser Pro Cys Asn 340 345 350 Gln His Ser Pro Tyr Trp Ala Pro Pro Cys Tyr Thr Leu Lys Pro Glu 355 360 365 Thr <210> 66 <211> 276 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 66 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Glu Val Gln Leu Val Glu Ser Gly Gly Gly 20 25 30 Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly 35 40 45 Phe Thr Phe Ser Ser Tyr Ala Met Ser Trp Val Arg Gln Ala Pro Gly 50 55 60 Lys Gly Leu Glu Trp Val Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr 65 70 75 80 Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn 85 90 95 Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp 100 105 110 Thr Ala Val Tyr Tyr Cys Ala Arg Gly Val Gly Ala Phe Arg Pro Tyr 115 120 125 Arg Lys His Glu Trp Gly Gln Gly Thr Leu Val Thr Val Ser Arg Gly 130 135 140 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Ser 145 150 155 160 Glu Leu Thr Gln Asp Pro Ala Val Ser Val Ala Leu Gly Gln Thr Val 165 170 175 Arg Ile Thr Cys Gln Gly Asp Ser Leu Arg Ser Tyr Tyr Ala Ser Trp 180 185 190 Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Ile Tyr Gly Lys 195 200 205 Asn Asn Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Ser Ser 210 215 220 Gly Asn Thr Ala Ser Leu Thr Thr Thr Gly Ala Gln Ala Glu Asp Glu 225 230 235 240 Ala Asp Tyr Tyr Cys Asn Ser Ser Pro Phe Glu His Asn Leu Val Val 245 250 255 Phe Gly Gly Gly Thr Lys Leu Thr Val Leu His His His His His His 260 265 270 Glu Pro Glu Ala 275 <210> 67 <211> 674 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 67 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr 20 25 30 Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn 35 40 45 Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe 50 55 60 Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys 65 70 75 80 Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln 85 90 95 Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn 100 105 110 Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu 115 120 125 Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile 130 135 140 Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly 145 150 155 160 Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val Glu 165 170 175 Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys 180 185 190 Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg 195 200 205 Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser 210 215 220 Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser 225 230 235 240 Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg 245 250 255 Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp 260 265 270 Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly 275 280 285 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile 290 295 300 Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg 305 310 315 320 Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly 325 330 335 Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser 340 345 350 Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly 355 360 365 Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp 370 375 380 Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe 385 390 395 400 Gly Gly Gly Thr Lys Val Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly 405 410 415 Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly 420 425 430 Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala 435 440 445 Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala 450 455 460 Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg 465 470 475 480 Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg 485 490 495 Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro 500 505 510 Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val 515 520 525 Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser 530 535 540 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Glu 545 550 555 560 Ser Gly Gly Gly Leu Val Gln Ala Gly Gly Ser Leu Arg Leu Ser Cys 565 570 575 Ala Ala Ser Gly Arg Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr Arg 580 585 590 Gln Ala Pro Gly Lys Gln Arg Glu Leu Val Ala Arg Ile Thr Arg Gly 595 600 605 Gly Thr Ile Ser Tyr Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser 610 615 620 Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys 625 630 635 640 Pro Glu Asp Thr Gly Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr Asp 645 650 655 Tyr Trp Gly Lys Gly Thr Gln Val Thr Val Ser Ser His His His His 660 665 670 His His <210> 68 <211> 674 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 68 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Glu Ser Gly Gly Gly 20 25 30 Leu Val Gln Ala Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly 35 40 45 Arg Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly 50 55 60 Lys Gln Arg Glu Leu Val Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser 65 70 75 80 Tyr Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala 85 90 95 Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr 100 105 110 Gly Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys 115 120 125 Gly Thr Gln Val Thr Val Ser Ser Ser Gly Gly Pro Gly Pro Ala Gly 130 135 140 Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly 145 150 155 160 Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser 165 170 175 Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro 180 185 190 Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp 195 200 205 Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp 210 215 220 Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu 225 230 235 240 Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser 245 250 255 Ser Gln Gly Thr Leu Val Thr Val Ser Ser Ser Gly Gly Pro Gly Pro 260 265 270 Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val Glu Ser 275 280 285 Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala 290 295 300 Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg Gln 305 310 315 320 Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser Ser 325 330 335 Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser Arg 340 345 350 Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala 355 360 365 Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp Ala 370 375 380 Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly 385 390 395 400 Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln 405 410 415 Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val 420 425 430 Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly Trp 435 440 445 Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser Ala 450 455 460 Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser 465 470 475 480 Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe 485 490 495 Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe Gly 500 505 510 Gly Gly Thr Lys Val Glu Ile Lys Ser Gly Gly Pro Gly Pro Ala Gly 515 520 525 Met Lys Gly Leu Pro Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys 530 535 540 Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu 545 550 555 560 Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr 565 570 575 Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln 580 585 590 Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala 595 600 605 Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile 610 615 620 Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys 625 630 635 640 Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp 645 650 655 Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr His His His His 660 665 670 His His <210> 69 <211> 425 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 69 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile 20 25 30 Pro His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu Gly Thr Met Leu 35 40 45 Asn Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu 50 55 60 Tyr Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser Trp Asp Asn Gln 65 70 75 80 Cys Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr Lys Gln Val Thr 85 90 95 Pro Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr Glu Met Gln Ser 100 105 110 Pro Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly His Cys Arg Glu 115 120 125 Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile Tyr His Phe Val 130 135 140 Val Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu 145 150 155 160 His Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr His Gly Lys Thr 165 170 175 Arg Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly Glu Met Glu Thr Ser 180 185 190 Gln Phe Pro Gly Glu Glu Lys Pro Gln Ala Ser Pro Glu Gly Arg Pro 195 200 205 Glu Ser Glu Thr Ser Cys Leu Val Thr Thr Thr Asp Phe Gln Ile Gln 210 215 220 Thr Glu Met Ala Ala Thr Met Glu Thr Ser Ile Phe Thr Thr Glu Tyr 225 230 235 240 Gln Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 245 250 255 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser 260 265 270 Gly Gly Pro Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Ala Pro 275 280 285 Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu 290 295 300 Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro 305 310 315 320 Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala 325 330 335 Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu 340 345 350 Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro 355 360 365 Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly 370 375 380 Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile 385 390 395 400 Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile Ile Ser 405 410 415 Thr Leu Thr His His His His His His 420 425 <210> 70 <211> 425 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 70 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr 20 25 30 Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn 35 40 45 Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe 50 55 60 Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys 65 70 75 80 Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln 85 90 95 Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn 100 105 110 Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu 115 120 125 Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile 130 135 140 Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly 145 150 155 160 Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Gly Gly Gly Gly Ser Gly 165 170 175 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 180 185 190 Gly Gly Ser Gly Gly Gly Gly Ser Glu Leu Cys Asp Asp Asp Pro Pro 195 200 205 Glu Ile Pro His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu Gly Thr 210 215 220 Met Leu Asn Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile Lys Ser Gly 225 230 235 240 Ser Leu Tyr Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser Trp Asp 245 250 255 Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr Lys Gln 260 265 270 Val Thr Pro Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr Glu Met 275 280 285 Gln Ser Pro Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly His Cys 290 295 300 Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile Tyr His 305 310 315 320 Phe Val Val Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly Tyr Arg 325 330 335 Ala Leu His Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr His Gly 340 345 350 Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly Glu Met Glu 355 360 365 Thr Ser Gln Phe Pro Gly Glu Glu Lys Pro Gln Ala Ser Pro Glu Gly 370 375 380 Arg Pro Glu Ser Glu Thr Ser Cys Leu Val Thr Thr Thr Asp Phe Gln 385 390 395 400 Ile Gln Thr Glu Met Ala Ala Thr Met Glu Thr Ser Ile Phe Thr Thr 405 410 415 Glu Tyr Gln His His His His His His 420 425 <210> 71 <211> 555 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 71 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Glu Val Gln Leu Val Glu Ser Gly Gly Gly 20 25 30 Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly 35 40 45 Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly 50 55 60 Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr 65 70 75 80 Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn 85 90 95 Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp 100 105 110 Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser 115 120 125 Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly 130 135 140 Gly Gly Ser Gly Gly Gly Gly Ser Glu Leu Cys Asp Asp Asp Pro Pro 145 150 155 160 Glu Ile Pro His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu Gly Thr 165 170 175 Met Leu Asn Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile Lys Ser Gly 180 185 190 Ser Leu Tyr Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser Trp Asp 195 200 205 Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr Lys Gln 210 215 220 Val Thr Pro Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr Glu Met 225 230 235 240 Gln Ser Pro Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly His Cys 245 250 255 Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile Tyr His 260 265 270 Phe Val Val Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly Tyr Arg 275 280 285 Ala Leu His Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr His Gly 290 295 300 Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly Glu Met Glu 305 310 315 320 Thr Ser Gln Phe Pro Gly Glu Glu Lys Pro Gln Ala Ser Pro Glu Gly 325 330 335 Arg Pro Glu Ser Glu Thr Ser Cys Leu Val Thr Thr Thr Asp Phe Gln 340 345 350 Ile Gln Thr Glu Met Ala Ala Thr Met Glu Thr Ser Ile Phe Thr Thr 355 360 365 Glu Tyr Gln Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 370 375 380 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 385 390 395 400 Ser Ser Gly Gly Pro Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser 405 410 415 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 420 425 430 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 435 440 445 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 450 455 460 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 465 470 475 480 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 485 490 495 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 500 505 510 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 515 520 525 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile 530 535 540 Ile Ser Thr Leu Thr His His His His His His 545 550 555 <210> 72 <211> 555 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 72 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr 20 25 30 Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn 35 40 45 Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe 50 55 60 Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys 65 70 75 80 Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln 85 90 95 Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn 100 105 110 Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu 115 120 125 Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile 130 135 140 Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly 145 150 155 160 Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Gly Gly Gly Gly Ser Gly 165 170 175 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 180 185 190 Gly Gly Ser Gly Gly Gly Gly Ser Glu Leu Cys Asp Asp Asp Pro Pro 195 200 205 Glu Ile Pro His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu Gly Thr 210 215 220 Met Leu Asn Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile Lys Ser Gly 225 230 235 240 Ser Leu Tyr Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser Trp Asp 245 250 255 Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr Lys Gln 260 265 270 Val Thr Pro Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr Glu Met 275 280 285 Gln Ser Pro Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly His Cys 290 295 300 Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile Tyr His 305 310 315 320 Phe Val Val Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly Tyr Arg 325 330 335 Ala Leu His Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr His Gly 340 345 350 Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly Glu Met Glu 355 360 365 Thr Ser Gln Phe Pro Gly Glu Glu Lys Pro Gln Ala Ser Pro Glu Gly 370 375 380 Arg Pro Glu Ser Glu Thr Ser Cys Leu Val Thr Thr Thr Asp Phe Gln 385 390 395 400 Ile Gln Thr Glu Met Ala Ala Thr Met Glu Thr Ser Ile Phe Thr Thr 405 410 415 Glu Tyr Gln Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 420 425 430 Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro 435 440 445 Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser 450 455 460 Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu 465 470 475 480 Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr Leu Tyr Ala Glu 485 490 495 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Thr Thr 500 505 510 Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr 515 520 525 Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser Gln Gly Thr Leu 530 535 540 Val Thr Val Ser Ser His His His His His His 545 550 555 <210> 73 <211> 555 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 73 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr 20 25 30 Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn 35 40 45 Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe 50 55 60 Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys 65 70 75 80 Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln 85 90 95 Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn 100 105 110 Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu 115 120 125 Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile 130 135 140 Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly 145 150 155 160 Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Gly Gly Gly Gly Ser Gly 165 170 175 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 180 185 190 Gly Gly Ser Gly Gly Gly Gly Ser Glu Leu Cys Asp Asp Asp Pro Pro 195 200 205 Glu Ile Pro His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu Gly Thr 210 215 220 Met Leu Asn Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile Lys Ser Gly 225 230 235 240 Ser Leu Tyr Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser Trp Asp 245 250 255 Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr Lys Gln 260 265 270 Val Thr Pro Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr Glu Met 275 280 285 Gln Ser Pro Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly His Cys 290 295 300 Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile Tyr His 305 310 315 320 Phe Val Val Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly Tyr Arg 325 330 335 Ala Leu His Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr His Gly 340 345 350 Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly Glu Met Glu 355 360 365 Thr Ser Gln Phe Pro Gly Glu Glu Lys Pro Gln Ala Ser Pro Glu Gly 370 375 380 Arg Pro Glu Ser Glu Thr Ser Cys Leu Val Thr Thr Thr Asp Phe Gln 385 390 395 400 Ile Gln Thr Glu Met Ala Ala Thr Met Glu Thr Ser Ile Phe Thr Thr 405 410 415 Glu Tyr Gln Ser Gly Gly Pro Gly Pro Ala Gly Met Lys Gly Leu Pro 420 425 430 Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro 435 440 445 Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser 450 455 460 Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu 465 470 475 480 Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr Leu Tyr Ala Glu 485 490 495 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Thr Thr 500 505 510 Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr 515 520 525 Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser Gln Gly Thr Leu 530 535 540 Val Thr Val Ser Ser His His His His His His 545 550 555 <210> 74 <211> 575 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 74 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Glu Val Gln Leu Val Glu Ser Gly Gly Gly 20 25 30 Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly 35 40 45 Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly 50 55 60 Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr 65 70 75 80 Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn 85 90 95 Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp 100 105 110 Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser 115 120 125 Gln Gly Thr Leu Val Thr Val Ser Ser Ser Gly Gly Pro Gly Pro Ala 130 135 140 Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val Glu Ser Gly 145 150 155 160 Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala 165 170 175 Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg Gln Ala 180 185 190 Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser Ser Tyr 195 200 205 Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser Arg Asp 210 215 220 Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu 225 230 235 240 Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp Ala Leu 245 250 255 Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly 260 265 270 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met 275 280 285 Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr 290 295 300 Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly Trp Tyr 305 310 315 320 Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser Ala Ser 325 330 335 Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly 340 345 350 Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala 355 360 365 Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe Gly Gly 370 375 380 Gly Thr Lys Val Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly 385 390 395 400 Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 405 410 415 Gly Gly Gly Gly Ser Ser Gly Gly Pro Gly Pro Ala Gly Met Lys Gly 420 425 430 Leu Pro Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu 435 440 445 Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile 450 455 460 Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe 465 470 475 480 Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu 485 490 495 Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys 500 505 510 Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile 515 520 525 Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala 530 535 540 Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe 545 550 555 560 Cys Gln Ser Ile Ile Ser Thr Leu Thr His His His His His His 565 570 575 <210> 75 <211> 704 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 75 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr 20 25 30 Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn 35 40 45 Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe 50 55 60 Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys 65 70 75 80 Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln 85 90 95 Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn 100 105 110 Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu 115 120 125 Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile 130 135 140 Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly 145 150 155 160 Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Gly Gly Gly Gly Ser Gly 165 170 175 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 180 185 190 Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly 195 200 205 Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala 210 215 220 Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg Gln Ala 225 230 235 240 Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser Ser Tyr 245 250 255 Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser Arg Asp 260 265 270 Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu 275 280 285 Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp Ala Leu 290 295 300 Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ser Gly Gly 305 310 315 320 Pro Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Asp Ile Gln Met 325 330 335 Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr 340 345 350 Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly Trp Tyr 355 360 365 Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser Ala Ser 370 375 380 Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly 385 390 395 400 Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala 405 410 415 Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe Gly Gly 420 425 430 Gly Thr Lys Val Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly 435 440 445 Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly 450 455 460 Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly 465 470 475 480 Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly 485 490 495 Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr 500 505 510 Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn 515 520 525 Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp 530 535 540 Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser 545 550 555 560 Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly 565 570 575 Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Glu Ser Gly 580 585 590 Gly Gly Leu Val Gln Ala Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala 595 600 605 Ser Gly Arg Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala 610 615 620 Pro Gly Lys Gln Arg Glu Leu Val Ala Arg Ile Thr Arg Gly Gly Thr 625 630 635 640 Ile Ser Tyr Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp 645 650 655 Asn Ala Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu 660 665 670 Asp Thr Gly Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp 675 680 685 Gly Lys Gly Thr Gln Val Thr Val Ser Ser His His His His His His 690 695 700 <210> 76 <211> 689 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 76 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Glu Ser Gly Gly Gly 20 25 30 Leu Val Gln Ala Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly 35 40 45 Arg Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly 50 55 60 Lys Gln Arg Glu Leu Val Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser 65 70 75 80 Tyr Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala 85 90 95 Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr 100 105 110 Gly Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys 115 120 125 Gly Thr Gln Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly 130 135 140 Gly Ser Gly Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys 145 150 155 160 Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu 165 170 175 Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr 180 185 190 Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln 195 200 205 Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala 210 215 220 Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile 225 230 235 240 Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys 245 250 255 Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp 260 265 270 Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro 275 280 285 Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val 290 295 300 Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser 305 310 315 320 Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val 325 330 335 Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly 340 345 350 Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr 355 360 365 Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser 370 375 380 Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser 385 390 395 400 Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly 405 410 415 Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 420 425 430 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu 435 440 445 Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu 450 455 460 Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp 465 470 475 480 Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp 485 490 495 Ser Ser Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr 500 505 510 Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser 515 520 525 Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn 530 535 540 Trp Asp Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser 545 550 555 560 Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 565 570 575 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 580 585 590 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn 595 600 605 Val Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile 610 615 620 Tyr Ser Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly 625 630 635 640 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 645 650 655 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr 660 665 670 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys His His His His His 675 680 685 His <210> 77 <211> 545 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 77 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr 20 25 30 Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn 35 40 45 Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe 50 55 60 Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys 65 70 75 80 Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln 85 90 95 Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn 100 105 110 Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu 115 120 125 Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile 130 135 140 Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly 145 150 155 160 Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val Glu 165 170 175 Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys 180 185 190 Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg 195 200 205 Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser 210 215 220 Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser 225 230 235 240 Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg 245 250 255 Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp 260 265 270 Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly 275 280 285 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile 290 295 300 Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg 305 310 315 320 Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly 325 330 335 Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser 340 345 350 Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly 355 360 365 Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp 370 375 380 Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe 385 390 395 400 Gly Gly Gly Thr Lys Val Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly 405 410 415 Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly 420 425 430 Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala 435 440 445 Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala 450 455 460 Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg 465 470 475 480 Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg 485 490 495 Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro 500 505 510 Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val 515 520 525 Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser His His His His His 530 535 540 His 545 <210> 78 <211> 575 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 78 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr 20 25 30 Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn 35 40 45 Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe 50 55 60 Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys 65 70 75 80 Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln 85 90 95 Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn 100 105 110 Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu 115 120 125 Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile 130 135 140 Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly 145 150 155 160 Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Gly Gly Gly Gly Ser Gly 165 170 175 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 180 185 190 Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly 195 200 205 Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala 210 215 220 Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg Gln Ala 225 230 235 240 Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser Ser Tyr 245 250 255 Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser Arg Asp 260 265 270 Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu 275 280 285 Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp Ala Leu 290 295 300 Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly 305 310 315 320 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met 325 330 335 Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr 340 345 350 Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly Trp Tyr 355 360 365 Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser Ala Ser 370 375 380 Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly 385 390 395 400 Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala 405 410 415 Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe Gly Gly 420 425 430 Gly Thr Lys Val Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly 435 440 445 Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly 450 455 460 Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly 465 470 475 480 Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly 485 490 495 Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr 500 505 510 Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn 515 520 525 Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp 530 535 540 Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser 545 550 555 560 Gln Gly Thr Leu Val Thr Val Ser Ser His His His His His His 565 570 575 <210> 79 <211> 421 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 79 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Glu Val Gln Leu Val Glu Ser Gly Gly Gly 20 25 30 Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly 35 40 45 Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly 50 55 60 Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr 65 70 75 80 Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn 85 90 95 Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp 100 105 110 Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser 115 120 125 Gln Gly Thr Leu Val Thr Val Ser Ser Ser Gly Gly Pro Gly Pro Ala 130 135 140 Gly Met Lys Gly Leu Pro Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys 145 150 155 160 Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile 165 170 175 Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu 180 185 190 Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu 195 200 205 Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu 210 215 220 Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn 225 230 235 240 Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met 245 250 255 Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg 260 265 270 Trp Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly 275 280 285 Pro Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu 290 295 300 Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu 305 310 315 320 Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp 325 330 335 Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser 340 345 350 Gly Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe 355 360 365 Thr Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn 370 375 380 Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly 385 390 395 400 Ser Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser His 405 410 415 His His His His His 420 <210> 80 <211> 806 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 80 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Glu Ser Gly Gly Gly 20 25 30 Leu Val Gln Ala Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly 35 40 45 Arg Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly 50 55 60 Lys Gln Arg Glu Leu Val Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser 65 70 75 80 Tyr Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala 85 90 95 Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr 100 105 110 Gly Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys 115 120 125 Gly Thr Gln Val Thr Val Ser Ser Gly Ser Gly Ser Gly Ser Gly Ser 130 135 140 Gly Ser Gly Ser Gly Ser Gly Ser Glu Val Gln Leu Val Glu Ser Gly 145 150 155 160 Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala 165 170 175 Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala 180 185 190 Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg 195 200 205 Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg 210 215 220 Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro 225 230 235 240 Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val 245 250 255 Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Ser Gly Ser Gly 260 265 270 Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gln Val Gln Leu Gln 275 280 285 Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly Ser Leu Arg Leu Ser 290 295 300 Cys Ala Ala Ser Gly Arg Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr 305 310 315 320 Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val Ala Arg Ile Thr Arg 325 330 335 Gly Gly Thr Ile Ser Tyr Asp Asp Ser Val Lys Gly Arg Phe Thr Ile 340 345 350 Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu 355 360 365 Lys Pro Glu Asp Thr Gly Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr 370 375 380 Asp Tyr Trp Gly Lys Gly Thr Gln Val Thr Val Ser Ser Gly Ser Gly 385 390 395 400 Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Glu Val Gln 405 410 415 Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg 420 425 430 Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala 435 440 445 Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile 450 455 460 Asp Ser Ser Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe 465 470 475 480 Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn 485 490 495 Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser 500 505 510 Asn Trp Asp Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val 515 520 525 Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 530 535 540 Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val 545 550 555 560 Gly Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr 565 570 575 Asn Val Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu 580 585 590 Ile Tyr Ser Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser 595 600 605 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln 610 615 620 Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro 625 630 635 640 Tyr Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Ser Gly Gly Pro 645 650 655 Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Ala Pro Thr Ser Ser 660 665 670 Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu 675 680 685 Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr 690 695 700 Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu 705 710 715 720 Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val 725 730 735 Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu 740 745 750 Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr 755 760 765 Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe 770 775 780 Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr 785 790 795 800 His His His His His His 805 <210> 81 <211> 676 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 81 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Glu Ser Gly Gly Gly 20 25 30 Leu Val Gln Ala Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly 35 40 45 Arg Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly 50 55 60 Lys Gln Arg Glu Leu Val Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser 65 70 75 80 Tyr Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala 85 90 95 Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr 100 105 110 Gly Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys 115 120 125 Gly Thr Gln Val Thr Val Ser Ser Gly Ser Gly Ser Gly Ser Gly Ser 130 135 140 Gly Ser Gly Ser Gly Ser Gly Ser Glu Val Gln Leu Val Glu Ser Gly 145 150 155 160 Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala 165 170 175 Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala 180 185 190 Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg 195 200 205 Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg 210 215 220 Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro 225 230 235 240 Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val 245 250 255 Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Ser Gly Ser Gly 260 265 270 Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Glu Val Gln Leu Val 275 280 285 Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser 290 295 300 Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val 305 310 315 320 Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser 325 330 335 Ser Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile 340 345 350 Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu 355 360 365 Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp 370 375 380 Asp Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 385 390 395 400 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp 405 410 415 Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 420 425 430 Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val 435 440 445 Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr 450 455 460 Ser Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 465 470 475 480 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 485 490 495 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr 500 505 510 Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Ser Gly Gly Pro Gly Pro 515 520 525 Ala Gly Met Lys Gly Leu Pro Gly Ser Ala Pro Thr Ser Ser Ser Thr 530 535 540 Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met 545 550 555 560 Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met 565 570 575 Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His 580 585 590 Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn 595 600 605 Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser 610 615 620 Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe 625 630 635 640 Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn 645 650 655 Arg Trp Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr His His 660 665 670 His His His His 675 <210> 82 <211> 421 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 82 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Glu Ser Gly Gly Gly 20 25 30 Leu Val Gln Ala Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly 35 40 45 Arg Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly 50 55 60 Lys Gln Arg Glu Leu Val Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser 65 70 75 80 Tyr Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala 85 90 95 Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr 100 105 110 Gly Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys 115 120 125 Gly Thr Gln Val Thr Val Ser Ser Gly Ser Gly Ser Gly Ser Gly Ser 130 135 140 Gly Ser Gly Ser Gly Ser Gly Ser Glu Val Gln Leu Val Glu Ser Gly 145 150 155 160 Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala 165 170 175 Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala 180 185 190 Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg 195 200 205 Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg 210 215 220 Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro 225 230 235 240 Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val 245 250 255 Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Ser Gly Gly Pro Gly 260 265 270 Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Ala Pro Thr Ser Ser Ser 275 280 285 Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln 290 295 300 Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg 305 310 315 320 Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys 325 330 335 His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu 340 345 350 Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile 355 360 365 Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr 370 375 380 Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu 385 390 395 400 Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr His 405 410 415 His His His His His 420 <210> 83 <211> 420 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 83 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Glu Ser Gly Gly Gly 20 25 30 Leu Val Gln Ala Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly 35 40 45 Arg Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly 50 55 60 Lys Gln Arg Glu Leu Val Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser 65 70 75 80 Tyr Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala 85 90 95 Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr 100 105 110 Gly Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys 115 120 125 Gly Thr Gln Val Thr Val Ser Ser Ser Gly Gly Pro Gly Pro Ala Gly 130 135 140 Met Lys Gly Leu Pro Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys 145 150 155 160 Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu 165 170 175 Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr 180 185 190 Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln 195 200 205 Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala 210 215 220 Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile 225 230 235 240 Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys 245 250 255 Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp 260 265 270 Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro 275 280 285 Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val 290 295 300 Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser 305 310 315 320 Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val 325 330 335 Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly 340 345 350 Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr 355 360 365 Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser 370 375 380 Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser 385 390 395 400 Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser His His 405 410 415 His His His His 420 <210> 84 <211> 550 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 84 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Glu Ser Gly Gly Gly 20 25 30 Leu Val Gln Ala Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly 35 40 45 Arg Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly 50 55 60 Lys Gln Arg Glu Leu Val Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser 65 70 75 80 Tyr Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala 85 90 95 Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr 100 105 110 Gly Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys 115 120 125 Gly Thr Gln Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly 130 135 140 Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly 145 150 155 160 Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser 165 170 175 Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro 180 185 190 Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp 195 200 205 Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp 210 215 220 Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu 225 230 235 240 Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser 245 250 255 Ser Gln Gly Thr Leu Val Thr Val Ser Ser Ser Gly Gly Pro Gly Pro 260 265 270 Ala Gly Met Lys Gly Leu Pro Gly Ser Ala Pro Thr Ser Ser Ser Thr 275 280 285 Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met 290 295 300 Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met 305 310 315 320 Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His 325 330 335 Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn 340 345 350 Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser 355 360 365 Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe 370 375 380 Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn 385 390 395 400 Arg Trp Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly 405 410 415 Gly Pro Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln 420 425 430 Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg 435 440 445 Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser 450 455 460 Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile 465 470 475 480 Ser Gly Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg 485 490 495 Phe Thr Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met 500 505 510 Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly 515 520 525 Gly Ser Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser 530 535 540 His His His His His His 545 550 <210> 85 <211> 420 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 85 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Glu Ser Gly Gly Gly 20 25 30 Leu Val Gln Ala Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly 35 40 45 Arg Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly 50 55 60 Lys Gln Arg Glu Leu Val Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser 65 70 75 80 Tyr Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala 85 90 95 Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr 100 105 110 Gly Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys 115 120 125 Gly Thr Gln Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly 130 135 140 Gly Ser Gly Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys 145 150 155 160 Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu 165 170 175 Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr 180 185 190 Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln 195 200 205 Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala 210 215 220 Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile 225 230 235 240 Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys 245 250 255 Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp 260 265 270 Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro 275 280 285 Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val 290 295 300 Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser 305 310 315 320 Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val 325 330 335 Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly 340 345 350 Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr 355 360 365 Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser 370 375 380 Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser 385 390 395 400 Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser His His 405 410 415 His His His His 420 <210> 86 <211> 290 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 86 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Glu Ser Gly Gly Gly 20 25 30 Leu Val Gln Ala Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly 35 40 45 Arg Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly 50 55 60 Lys Gln Arg Glu Leu Val Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser 65 70 75 80 Tyr Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala 85 90 95 Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr 100 105 110 Gly Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys 115 120 125 Gly Thr Gln Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly 130 135 140 Gly Ser Gly Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys 145 150 155 160 Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu 165 170 175 Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr 180 185 190 Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln 195 200 205 Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala 210 215 220 Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile 225 230 235 240 Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys 245 250 255 Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp 260 265 270 Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr His His His His 275 280 285 His His 290 <210> 87 <211> 291 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 87 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr 20 25 30 Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn 35 40 45 Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe 50 55 60 Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys 65 70 75 80 Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln 85 90 95 Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn 100 105 110 Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu 115 120 125 Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile 130 135 140 Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly 145 150 155 160 Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val Glu 165 170 175 Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys 180 185 190 Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg 195 200 205 Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser 210 215 220 Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile 225 230 235 240 Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu 245 250 255 Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu 260 265 270 Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser His His His 275 280 285 His His His 290 <210> 88 <211> 689 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 88 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Glu Ser Gly Gly Gly 20 25 30 Leu Val Gln Ala Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly 35 40 45 Arg Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly 50 55 60 Lys Gln Arg Glu Leu Val Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser 65 70 75 80 Tyr Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala 85 90 95 Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr 100 105 110 Gly Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys 115 120 125 Gly Thr Gln Val Thr Val Ser Ser Ser Gly Gly Pro Gly Pro Ala Gly 130 135 140 Met Lys Gly Leu Pro Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys 145 150 155 160 Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu 165 170 175 Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr 180 185 190 Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln 195 200 205 Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala 210 215 220 Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile 225 230 235 240 Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys 245 250 255 Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp 260 265 270 Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro 275 280 285 Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val 290 295 300 Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser 305 310 315 320 Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val 325 330 335 Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly 340 345 350 Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr 355 360 365 Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser 370 375 380 Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser 385 390 395 400 Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly 405 410 415 Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 420 425 430 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu 435 440 445 Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu 450 455 460 Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp 465 470 475 480 Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp 485 490 495 Ser Ser Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr 500 505 510 Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser 515 520 525 Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn 530 535 540 Trp Asp Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser 545 550 555 560 Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 565 570 575 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 580 585 590 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn 595 600 605 Val Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile 610 615 620 Tyr Ser Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly 625 630 635 640 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 645 650 655 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr 660 665 670 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys His His His His His 675 680 685 His <210> 89 <211> 704 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 89 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr 20 25 30 Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn 35 40 45 Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe 50 55 60 Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys 65 70 75 80 Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln 85 90 95 Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn 100 105 110 Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu 115 120 125 Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile 130 135 140 Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly 145 150 155 160 Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Gly Gly Gly Gly Ser Gly 165 170 175 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 180 185 190 Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly 195 200 205 Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala 210 215 220 Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg Gln Ala 225 230 235 240 Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser Ser Tyr 245 250 255 Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser Arg Asp 260 265 270 Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu 275 280 285 Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp Ala Leu 290 295 300 Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly 305 310 315 320 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met 325 330 335 Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr 340 345 350 Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly Trp Tyr 355 360 365 Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser Ala Ser 370 375 380 Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly 385 390 395 400 Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala 405 410 415 Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe Gly Gly 420 425 430 Gly Thr Lys Val Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly 435 440 445 Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly 450 455 460 Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly 465 470 475 480 Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly 485 490 495 Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr 500 505 510 Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn 515 520 525 Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp 530 535 540 Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser 545 550 555 560 Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly 565 570 575 Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Glu Ser Gly 580 585 590 Gly Gly Leu Ala Gln Ala Gly Gly Ser Leu Ser Leu Ser Cys Ala Ala 595 600 605 Ser Gly Phe Thr Val Ser Asn Ser Val Met Ala Trp Tyr Arg Gln Thr 610 615 620 Pro Gly Lys Gln Arg Glu Phe Val Ala Ile Ile Asn Ser Val Gly Ser 625 630 635 640 Thr Asn Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp 645 650 655 Asn Ala Lys Asn Thr Val Tyr Leu Gln Met Asn Asn Leu Lys Pro Glu 660 665 670 Asp Thr Ala Val Tyr Val Cys Asn Arg Asn Phe Asp Arg Ile Tyr Trp 675 680 685 Gly Gln Gly Thr Gln Val Thr Val Ser Ser His His His His His His 690 695 700 <210> 90 <211> 674 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 90 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Glu Ser Gly Gly Gly 20 25 30 Leu Ala Gln Ala Gly Gly Ser Leu Ser Leu Ser Cys Ala Ala Ser Gly 35 40 45 Phe Thr Val Ser Asn Ser Val Met Ala Trp Tyr Arg Gln Thr Pro Gly 50 55 60 Lys Gln Arg Glu Phe Val Ala Ile Ile Asn Ser Val Gly Ser Thr Asn 65 70 75 80 Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala 85 90 95 Lys Asn Thr Val Tyr Leu Gln Met Asn Asn Leu Lys Pro Glu Asp Thr 100 105 110 Ala Val Tyr Val Cys Asn Arg Asn Phe Asp Arg Ile Tyr Trp Gly Gln 115 120 125 Gly Thr Gln Val Thr Val Ser Ser Ala Pro Thr Ser Ser Ser Thr Lys 130 135 140 Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile 145 150 155 160 Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu 165 170 175 Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu 180 185 190 Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu 195 200 205 Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn 210 215 220 Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met 225 230 235 240 Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg 245 250 255 Trp Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly 260 265 270 Pro Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu 275 280 285 Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu 290 295 300 Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp 305 310 315 320 Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser 325 330 335 Gly Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe 340 345 350 Thr Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn 355 360 365 Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly 370 375 380 Ser Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly 385 390 395 400 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 405 410 415 Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln 420 425 430 Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg 435 440 445 Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala 450 455 460 Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile 465 470 475 480 Asp Ser Ser Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe 485 490 495 Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn 500 505 510 Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser 515 520 525 Asn Trp Asp Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val 530 535 540 Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 545 550 555 560 Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val 565 570 575 Gly Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr 580 585 590 Asn Val Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu 595 600 605 Ile Tyr Ser Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser 610 615 620 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln 625 630 635 640 Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro 645 650 655 Tyr Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys His His His His 660 665 670 His His <210> 91 <211> 704 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 91 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Glu Val Gln Leu Val Glu Ser Gly Gly Gly 20 25 30 Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly 35 40 45 Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg Gln Ala Pro Gly 50 55 60 Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser Ser Tyr Thr Tyr 65 70 75 80 Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala 85 90 95 Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr 100 105 110 Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp Ala Leu Asp Tyr 115 120 125 Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser 130 135 140 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln 145 150 155 160 Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr 165 170 175 Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly Trp Tyr Gln Gln 180 185 190 Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser Ala Ser Phe Arg 195 200 205 Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp 210 215 220 Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr 225 230 235 240 Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe Gly Gly Gly Thr 245 250 255 Lys Val Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 260 265 270 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 275 280 285 Gly Gly Ser Ser Gly Gly Pro Gly Pro Ala Gly Met Lys Gly Leu Pro 290 295 300 Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu 305 310 315 320 Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn 325 330 335 Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met 340 345 350 Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu 355 360 365 Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe 370 375 380 His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu 385 390 395 400 Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu 405 410 415 Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln 420 425 430 Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly Pro Ala Gly Met 435 440 445 Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly 450 455 460 Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly 465 470 475 480 Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly 485 490 495 Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr 500 505 510 Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn 515 520 525 Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp 530 535 540 Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser 545 550 555 560 Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly 565 570 575 Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Glu Ser Gly 580 585 590 Gly Gly Leu Ala Gln Ala Gly Gly Ser Leu Ser Leu Ser Cys Ala Ala 595 600 605 Ser Gly Phe Thr Val Ser Asn Ser Val Met Ala Trp Tyr Arg Gln Thr 610 615 620 Pro Gly Lys Gln Arg Glu Phe Val Ala Ile Ile Asn Ser Val Gly Ser 625 630 635 640 Thr Asn Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp 645 650 655 Asn Ala Lys Asn Thr Val Tyr Leu Gln Met Asn Asn Leu Lys Pro Glu 660 665 670 Asp Thr Ala Val Tyr Val Cys Asn Arg Asn Phe Asp Arg Ile Tyr Trp 675 680 685 Gly Gln Gly Thr Gln Val Thr Val Ser Ser His His His His His His 690 695 700 <210> 92 <211> 704 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 92 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Glu Ser Gly Gly Gly 20 25 30 Leu Ala Gln Ala Gly Gly Ser Leu Ser Leu Ser Cys Ala Ala Ser Gly 35 40 45 Phe Thr Val Ser Asn Ser Val Met Ala Trp Tyr Arg Gln Thr Pro Gly 50 55 60 Lys Gln Arg Glu Phe Val Ala Ile Ile Asn Ser Val Gly Ser Thr Asn 65 70 75 80 Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala 85 90 95 Lys Asn Thr Val Tyr Leu Gln Met Asn Asn Leu Lys Pro Glu Asp Thr 100 105 110 Ala Val Tyr Val Cys Asn Arg Asn Phe Asp Arg Ile Tyr Trp Gly Gln 115 120 125 Gly Thr Gln Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly 130 135 140 Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly 145 150 155 160 Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser 165 170 175 Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro 180 185 190 Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp 195 200 205 Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp 210 215 220 Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu 225 230 235 240 Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser 245 250 255 Ser Gln Gly Thr Leu Val Thr Val Ser Ser Ser Gly Gly Pro Gly Pro 260 265 270 Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val Glu Ser 275 280 285 Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala 290 295 300 Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg Gln 305 310 315 320 Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser Ser 325 330 335 Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser Arg 340 345 350 Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala 355 360 365 Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp Ala 370 375 380 Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly 385 390 395 400 Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln 405 410 415 Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val 420 425 430 Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly Trp 435 440 445 Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser Ala 450 455 460 Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser 465 470 475 480 Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe 485 490 495 Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe Gly 500 505 510 Gly Gly Thr Lys Val Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly Gly 515 520 525 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 530 535 540 Ser Gly Gly Gly Gly Ser Ser Gly Gly Pro Gly Pro Ala Gly Met Lys 545 550 555 560 Gly Leu Pro Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln 565 570 575 Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly 580 585 590 Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys 595 600 605 Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu 610 615 620 Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser 625 630 635 640 Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val 645 650 655 Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr 660 665 670 Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr 675 680 685 Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr His His His His His His 690 695 700 <210> 93 <211> 689 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 93 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Glu Val Gln Leu Val Glu Ser Gly Gly Gly 20 25 30 Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly 35 40 45 Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg Gln Ala Pro Gly 50 55 60 Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser Ser Tyr Thr Tyr 65 70 75 80 Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala 85 90 95 Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr 100 105 110 Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp Ala Leu Asp Tyr 115 120 125 Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser 130 135 140 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln 145 150 155 160 Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr 165 170 175 Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly Trp Tyr Gln Gln 180 185 190 Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser Ala Ser Phe Arg 195 200 205 Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp 210 215 220 Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr 225 230 235 240 Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe Gly Gly Gly Thr 245 250 255 Lys Val Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 260 265 270 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 275 280 285 Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln 290 295 300 Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe 305 310 315 320 Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu 325 330 335 Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr Leu Tyr Ala 340 345 350 Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Thr 355 360 365 Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val 370 375 380 Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser Gln Gly Thr 385 390 395 400 Leu Val Thr Val Ser Ser Ser Gly Gly Pro Gly Pro Ala Gly Met Lys 405 410 415 Gly Leu Pro Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln 420 425 430 Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly 435 440 445 Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys 450 455 460 Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu 465 470 475 480 Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser 485 490 495 Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val 500 505 510 Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr 515 520 525 Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr 530 535 540 Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Gly Gly Gly Gly Ser Gly 545 550 555 560 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Glu Ser 565 570 575 Gly Gly Gly Leu Ala Gln Ala Gly Gly Ser Leu Ser Leu Ser Cys Ala 580 585 590 Ala Ser Gly Phe Thr Val Ser Asn Ser Val Met Ala Trp Tyr Arg Gln 595 600 605 Thr Pro Gly Lys Gln Arg Glu Phe Val Ala Ile Ile Asn Ser Val Gly 610 615 620 Ser Thr Asn Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg 625 630 635 640 Asp Asn Ala Lys Asn Thr Val Tyr Leu Gln Met Asn Asn Leu Lys Pro 645 650 655 Glu Asp Thr Ala Val Tyr Val Cys Asn Arg Asn Phe Asp Arg Ile Tyr 660 665 670 Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser His His His His His 675 680 685 His <210> 94 <211> 700 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 94 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Glu Ser Gly Gly Gly 20 25 30 Leu Val Gln Ala Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly 35 40 45 Arg Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly 50 55 60 Lys Gln Arg Glu Leu Val Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser 65 70 75 80 Tyr Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala 85 90 95 Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr 100 105 110 Gly Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys 115 120 125 Gly Thr Gln Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly 130 135 140 Gly Ser Gly Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys 145 150 155 160 Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu 165 170 175 Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr 180 185 190 Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln 195 200 205 Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala 210 215 220 Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile 225 230 235 240 Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys 245 250 255 Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp 260 265 270 Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro 275 280 285 Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Arg Gly Glu Thr Gly 290 295 300 Pro Ala Ala Pro Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly 305 310 315 320 Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly 325 330 335 Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly 340 345 350 Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr 355 360 365 Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn 370 375 380 Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp 385 390 395 400 Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser 405 410 415 Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly 420 425 430 Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 435 440 445 Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly 450 455 460 Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser 465 470 475 480 Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg Gln Ala Pro 485 490 495 Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser Ser Tyr Thr 500 505 510 Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser Arg Asp Asn 515 520 525 Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp 530 535 540 Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp Ala Leu Asp 545 550 555 560 Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly 565 570 575 Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr 580 585 590 Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile 595 600 605 Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly Trp Tyr Gln 610 615 620 Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser Ala Ser Phe 625 630 635 640 Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr 645 650 655 Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr 660 665 670 Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe Gly Gly Gly 675 680 685 Thr Lys Val Glu Ile Lys His His His His His His 690 695 700 <210> 95 <211> 276 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 95 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Glu Val Gln Leu Val Glu Ser Gly Gly Gly 20 25 30 Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly 35 40 45 Phe Thr Phe Ser Ser Tyr Ala Met Ser Trp Val Arg Gln Ala Pro Gly 50 55 60 Lys Gly Leu Glu Trp Val Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr 65 70 75 80 Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn 85 90 95 Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp 100 105 110 Thr Ala Val Tyr Tyr Cys Ala Arg Gly Val Gly Ala Phe Arg Pro Tyr 115 120 125 Arg Lys His Glu Trp Gly Gln Gly Thr Leu Val Thr Val Ser Arg Gly 130 135 140 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Ser 145 150 155 160 Glu Leu Thr Gln Asp Pro Ala Val Ser Val Ala Leu Gly Gln Thr Val 165 170 175 Arg Ile Thr Cys Gln Gly Asp Ser Leu Arg Ser Tyr Tyr Ala Ser Trp 180 185 190 Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Ile Tyr Gly Lys 195 200 205 Asn Asn Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Ser Ser 210 215 220 Gly Asn Thr Ala Ser Leu Thr Thr Thr Gly Ala Gln Ala Glu Asp Glu 225 230 235 240 Ala Asp Tyr Tyr Cys Asn Ser Ser Pro Phe Glu His Asn Leu Val Val 245 250 255 Phe Gly Gly Gly Thr Lys Leu Thr Val Leu His His His His His His 260 265 270 Glu Pro Glu Ala 275 <210> 96 <211> 279 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 96 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Gln Ser Gly Ala Glu 20 25 30 Leu Val Arg Pro Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly 35 40 45 Tyr Ala Phe Thr Asn Tyr Leu Ile Glu Trp Val Lys Gln Arg Pro Gly 50 55 60 Gln Gly Leu Glu Trp Ile Gly Val Ile Asn Pro Gly Ser Gly Gly Thr 65 70 75 80 Asn Tyr Asn Glu Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys 85 90 95 Ser Ser Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp 100 105 110 Ser Ala Val Tyr Phe Cys Ala Arg Trp Arg Gly Asp Gly Tyr Tyr Ala 115 120 125 Tyr Phe Asp Val Trp Gly Ala Gly Thr Thr Val Thr Val Ser Ser Gly 130 135 140 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile 145 150 155 160 Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly Gln Arg 165 170 175 Ala Thr Ile Ser Cys Lys Ala Ser Gln Ser Val Asp Tyr Asp Gly Asp 180 185 190 Ser Tyr Met Asn Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu 195 200 205 Leu Ile Tyr Ala Ala Ser Asn Leu Glu Ser Gly Ile Pro Ala Arg Phe 210 215 220 Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Asn Ile His Pro Val 225 230 235 240 Glu Glu Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Ser Asn Glu Asp 245 250 255 Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys His His His 260 265 270 His His His Glu Pro Glu Ala 275 <210> 97 <211> 279 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 97 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Gln Ser Gly Ala Glu 20 25 30 Leu Val Arg Pro Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly 35 40 45 Tyr Ala Phe Thr Asn Tyr Leu Ile Glu Trp Val Lys Gln Arg Pro Gly 50 55 60 Gln Gly Leu Glu Trp Ile Gly Val Ile Asn Pro Gly Ser Gly Gly Thr 65 70 75 80 Asn Tyr Asn Glu Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys 85 90 95 Ser Ser Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp 100 105 110 Ser Ala Val Tyr Phe Cys Ala Arg Trp Arg Gly Asp Gly Tyr Tyr Ala 115 120 125 Tyr Phe Asp Val Trp Gly Ala Gly Thr Thr Val Thr Val Ser Ser Ser 130 135 140 Gly Gly Pro Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Asp Ile 145 150 155 160 Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly Gln Arg 165 170 175 Ala Thr Ile Ser Cys Lys Ala Ser Gln Ser Val Asp Tyr Asp Gly Asp 180 185 190 Ser Tyr Met Asn Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu 195 200 205 Leu Ile Tyr Ala Ala Ser Asn Leu Glu Ser Gly Ile Pro Ala Arg Phe 210 215 220 Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Asn Ile His Pro Val 225 230 235 240 Glu Glu Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Ser Asn Glu Asp 245 250 255 Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys His His His 260 265 270 His His His Glu Pro Glu Ala 275 <210> 98 <211> 279 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 98 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Asp Ile Val Leu Thr Gln Ser Pro Ala Ser 20 25 30 Leu Ala Val Ser Leu Gly Gln Arg Ala Thr Ile Ser Cys Lys Ala Ser 35 40 45 Gln Ser Val Asp Tyr Asp Gly Asp Ser Tyr Met Asn Trp Tyr Gln Gln 50 55 60 Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Ala Ala Ser Asn Leu 65 70 75 80 Glu Ser Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp 85 90 95 Phe Thr Leu Asn Ile His Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr 100 105 110 Tyr Cys Gln Gln Ser Asn Glu Asp Pro Tyr Thr Phe Gly Gly Gly Thr 115 120 125 Lys Leu Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 130 135 140 Gly Gly Gly Ser Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val 145 150 155 160 Arg Pro Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala 165 170 175 Phe Thr Asn Tyr Leu Ile Glu Trp Val Lys Gln Arg Pro Gly Gln Gly 180 185 190 Leu Glu Trp Ile Gly Val Ile Asn Pro Gly Ser Gly Gly Thr Asn Tyr 195 200 205 Asn Glu Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser 210 215 220 Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala 225 230 235 240 Val Tyr Phe Cys Ala Arg Trp Arg Gly Asp Gly Tyr Tyr Ala Tyr Phe 245 250 255 Asp Val Trp Gly Ala Gly Thr Thr Val Thr Val Ser Ser His His His 260 265 270 His His His Glu Pro Glu Ala 275 <210> 99 <211> 279 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 99 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Asp Ile Val Leu Thr Gln Ser Pro Ala Ser 20 25 30 Leu Ala Val Ser Leu Gly Gln Arg Ala Thr Ile Ser Cys Lys Ala Ser 35 40 45 Gln Ser Val Asp Tyr Asp Gly Asp Ser Tyr Met Asn Trp Tyr Gln Gln 50 55 60 Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Ala Ala Ser Asn Leu 65 70 75 80 Glu Ser Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp 85 90 95 Phe Thr Leu Asn Ile His Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr 100 105 110 Tyr Cys Gln Gln Ser Asn Glu Asp Pro Tyr Thr Phe Gly Gly Gly Thr 115 120 125 Lys Leu Glu Ile Lys Ser Gly Gly Pro Gly Pro Ala Gly Met Lys Gly 130 135 140 Leu Pro Gly Ser Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val 145 150 155 160 Arg Pro Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala 165 170 175 Phe Thr Asn Tyr Leu Ile Glu Trp Val Lys Gln Arg Pro Gly Gln Gly 180 185 190 Leu Glu Trp Ile Gly Val Ile Asn Pro Gly Ser Gly Gly Thr Asn Tyr 195 200 205 Asn Glu Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser 210 215 220 Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala 225 230 235 240 Val Tyr Phe Cys Ala Arg Trp Arg Gly Asp Gly Tyr Tyr Ala Tyr Phe 245 250 255 Asp Val Trp Gly Ala Gly Thr Thr Val Thr Val Ser Ser His His His 260 265 270 His His His Glu Pro Glu Ala 275 <210> 100 <211> 295 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 100 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr 20 25 30 Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn 35 40 45 Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe 50 55 60 Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys 65 70 75 80 Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln 85 90 95 Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn 100 105 110 Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu 115 120 125 Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile 130 135 140 Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Gly Gly Gly Gly Ser 145 150 155 160 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu 165 170 175 Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys 180 185 190 Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg 195 200 205 Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser 210 215 220 Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile 225 230 235 240 Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu 245 250 255 Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu 260 265 270 Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser His His His 275 280 285 His His His Glu Pro Glu Ala 290 295 <210> 101 <211> 765 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 101 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr 20 25 30 Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn 35 40 45 Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe 50 55 60 Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys 65 70 75 80 Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln 85 90 95 Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn 100 105 110 Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu 115 120 125 Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile 130 135 140 Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Gly Gly Gly Gly Ser 145 150 155 160 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ala His Lys Ser Glu 165 170 175 Val Ala His Arg Phe Lys Asp Leu Gly Glu Glu Asn Phe Lys Ala Leu 180 185 190 Val Leu Ile Ala Phe Ala Gln Tyr Leu Gln Gln Cys Pro Phe Glu Asp 195 200 205 His Val Lys Leu Val Asn Glu Val Thr Glu Phe Ala Lys Thr Cys Val 210 215 220 Ala Asp Glu Ser Ala Glu Asn Cys Asp Lys Ser Leu His Thr Leu Phe 225 230 235 240 Gly Asp Lys Leu Cys Thr Val Ala Thr Leu Arg Glu Thr Tyr Gly Glu 245 250 255 Met Ala Asp Cys Cys Ala Lys Gln Glu Pro Glu Arg Asn Glu Cys Phe 260 265 270 Leu Gln His Lys Asp Asp Asn Pro Asn Leu Pro Arg Leu Val Arg Pro 275 280 285 Glu Val Asp Val Met Cys Thr Ala Phe His Asp Asn Glu Glu Thr Phe 290 295 300 Leu Lys Lys Tyr Leu Tyr Glu Ile Ala Arg Arg His Pro Tyr Phe Tyr 305 310 315 320 Ala Pro Glu Leu Leu Phe Phe Ala Lys Arg Tyr Lys Ala Ala Phe Thr 325 330 335 Glu Cys Cys Gln Ala Ala Asp Lys Ala Ala Cys Leu Leu Pro Lys Leu 340 345 350 Asp Glu Leu Arg Asp Glu Gly Lys Ala Ser Ser Ala Lys Gln Arg Leu 355 360 365 Lys Cys Ala Ser Leu Gln Lys Phe Gly Glu Arg Ala Phe Lys Ala Trp 370 375 380 Ala Val Ala Arg Leu Ser Gln Arg Phe Pro Lys Ala Glu Phe Ala Glu 385 390 395 400 Val Ser Lys Leu Val Thr Asp Leu Thr Lys Val His Thr Glu Cys Cys 405 410 415 His Gly Asp Leu Leu Glu Cys Ala Asp Asp Arg Ala Asp Leu Ala Lys 420 425 430 Tyr Ile Cys Glu Asn Gln Asp Ser Ile Ser Ser Lys Leu Lys Glu Cys 435 440 445 Cys Glu Lys Pro Leu Leu Glu Lys Ser His Cys Ile Ala Glu Val Glu 450 455 460 Asn Asp Glu Met Pro Ala Asp Leu Pro Ser Leu Ala Ala Asp Phe Val 465 470 475 480 Glu Ser Lys Asp Val Cys Lys Asn Tyr Ala Glu Ala Lys Asp Val Phe 485 490 495 Leu Gly Met Phe Leu Tyr Glu Tyr Ala Arg Arg His Pro Asp Tyr Ser 500 505 510 Val Val Leu Leu Leu Arg Leu Ala Lys Thr Tyr Glu Thr Thr Leu Glu 515 520 525 Lys Cys Cys Ala Ala Ala Asp Pro His Glu Cys Tyr Ala Lys Val Phe 530 535 540 Asp Glu Phe Lys Pro Leu Val Glu Glu Pro Gln Asn Leu Ile Lys Gln 545 550 555 560 Asn Cys Glu Leu Phe Glu Gln Leu Gly Glu Tyr Lys Phe Gln Asn Ala 565 570 575 Leu Leu Val Arg Tyr Thr Lys Lys Val Pro Gln Val Ser Thr Pro Thr 580 585 590 Leu Val Glu Val Ser Arg Asn Leu Gly Lys Val Gly Ser Lys Cys Cys 595 600 605 Lys His Pro Glu Ala Lys Arg Met Pro Cys Ala Glu Asp Tyr Leu Ser 610 615 620 Val Val Leu Asn Gln Leu Cys Val Leu His Glu Lys Thr Pro Val Ser 625 630 635 640 Asp Arg Val Thr Lys Cys Cys Thr Glu Ser Leu Val Asn Arg Arg Pro 645 650 655 Cys Phe Ser Ala Leu Glu Val Asp Glu Thr Tyr Val Pro Lys Glu Phe 660 665 670 Asn Ala Glu Thr Phe Thr Phe His Ala Asp Ile Cys Thr Leu Ser Glu 675 680 685 Lys Glu Arg Gln Ile Lys Lys Gln Thr Ala Leu Val Glu Leu Val Lys 690 695 700 His Lys Pro Lys Ala Thr Lys Glu Gln Leu Lys Ala Val Met Asp Asp 705 710 715 720 Phe Ala Ala Phe Val Glu Lys Cys Cys Lys Ala Asp Asp Lys Glu Thr 725 730 735 Cys Phe Ala Glu Glu Gly Lys Lys Leu Val Ala Ala Ser Gln Ala Ala 740 745 750 Leu Gly Leu His His His His His His Glu Pro Glu Ala 755 760 765 <210> 102 <211> 765 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 102 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr 20 25 30 Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn 35 40 45 Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe 50 55 60 Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys 65 70 75 80 Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln 85 90 95 Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn 100 105 110 Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu 115 120 125 Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile 130 135 140 Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Gly Gly Gly Gly Ser 145 150 155 160 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ala His Lys Ser Glu 165 170 175 Val Ala His Arg Phe Lys Asp Leu Gly Glu Glu Asn Phe Lys Ala Leu 180 185 190 Val Leu Ile Ala Phe Ala Gln Tyr Leu Gln Gln Cys Pro Phe Glu Asp 195 200 205 His Val Lys Leu Val Asn Glu Val Thr Glu Phe Ala Lys Thr Cys Val 210 215 220 Ala Asp Glu Ser Ala Glu Asn Cys Asp Lys Ser Leu His Thr Leu Phe 225 230 235 240 Gly Asp Lys Leu Cys Thr Val Ala Thr Leu Arg Glu Thr Tyr Gly Glu 245 250 255 Met Ala Asp Cys Cys Ala Lys Gln Glu Pro Glu Arg Asn Glu Cys Phe 260 265 270 Leu Gln His Lys Asp Asp Asn Pro Asn Leu Pro Arg Leu Val Arg Pro 275 280 285 Glu Val Asp Val Met Cys Thr Ala Phe His Asp Asn Glu Glu Thr Phe 290 295 300 Leu Lys Lys Tyr Leu Tyr Glu Ile Ala Arg Arg His Pro Tyr Phe Tyr 305 310 315 320 Ala Pro Glu Leu Leu Phe Phe Ala Lys Arg Tyr Lys Ala Ala Phe Thr 325 330 335 Glu Cys Cys Gln Ala Ala Asp Lys Ala Ala Cys Leu Leu Pro Lys Leu 340 345 350 Asp Glu Leu Arg Asp Glu Gly Lys Ala Ser Ser Ala Lys Gln Arg Leu 355 360 365 Lys Cys Ala Ser Leu Gln Lys Phe Gly Glu Arg Ala Phe Lys Ala Trp 370 375 380 Ala Val Ala Arg Leu Ser Gln Arg Phe Pro Lys Ala Glu Phe Ala Glu 385 390 395 400 Val Ser Lys Leu Val Thr Asp Leu Thr Lys Val His Thr Glu Cys Cys 405 410 415 His Gly Asp Leu Leu Glu Cys Ala Asp Asp Arg Ala Asp Leu Ala Lys 420 425 430 Tyr Ile Cys Glu Asn Gln Asp Ser Ile Ser Ser Lys Leu Lys Glu Cys 435 440 445 Cys Glu Lys Pro Leu Leu Glu Lys Ser His Cys Ile Ala Glu Val Glu 450 455 460 Asn Asp Glu Met Pro Ala Asp Leu Pro Ser Leu Ala Ala Asp Phe Val 465 470 475 480 Glu Ser Lys Asp Val Cys Lys Asn Tyr Ala Glu Ala Lys Asp Val Phe 485 490 495 Leu Gly Met Phe Leu Tyr Glu Tyr Ala Arg Arg His Pro Asp Tyr Ser 500 505 510 Val Val Leu Leu Leu Arg Leu Ala Lys Thr Tyr Glu Thr Thr Leu Glu 515 520 525 Lys Cys Cys Ala Ala Ala Asp Pro His Glu Cys Tyr Ala Lys Val Phe 530 535 540 Asp Glu Phe Lys Pro Leu Val Glu Glu Pro Gln Asn Leu Ile Lys Gln 545 550 555 560 Asn Cys Glu Leu Phe Glu Gln Leu Gly Glu Tyr Lys Phe Gln Asn Ala 565 570 575 Leu Leu Val Arg Tyr Thr Lys Lys Val Pro Gln Val Ser Thr Pro Thr 580 585 590 Leu Val Glu Val Ser Arg Asn Leu Gly Lys Val Gly Ser Lys Cys Cys 595 600 605 Lys His Pro Glu Ala Lys Arg Met Pro Cys Ala Glu Asp Tyr Leu Ser 610 615 620 Val Val Leu Asn Gln Leu Cys Val Leu His Glu Lys Thr Pro Val Ser 625 630 635 640 Asp Arg Val Thr Lys Cys Cys Thr Glu Ser Leu Val Asn Arg Arg Pro 645 650 655 Cys Phe Ser Ala Leu Glu Val Asp Glu Thr Tyr Val Pro Lys Glu Phe 660 665 670 Asn Ala Glu Thr Phe Thr Phe His Ala Asp Ile Cys Thr Leu Ser Glu 675 680 685 Lys Glu Arg Gln Ile Lys Lys Gln Thr Ala Leu Val Glu Leu Val Lys 690 695 700 His Lys Pro Lys Ala Thr Lys Glu Gln Leu Lys Ala Val Met Asp Asp 705 710 715 720 Phe Ala Ala Phe Val Glu Lys Cys Cys Lys Ala Asp Asp Lys Glu Thr 725 730 735 Cys Phe Ala Glu Glu Gly Lys Lys Leu Val Ala Ala Ser Gln Ala Ala 740 745 750 Leu Gly Leu His His His His His His Glu Pro Glu Ala 755 760 765 <210> 103 <211> 765 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 103 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr 20 25 30 Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn 35 40 45 Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe 50 55 60 Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys 65 70 75 80 Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln 85 90 95 Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn 100 105 110 Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu 115 120 125 Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile 130 135 140 Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly 145 150 155 160 Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Asp Ala His Lys Ser Glu 165 170 175 Val Ala His Arg Phe Lys Asp Leu Gly Glu Glu Asn Phe Lys Ala Leu 180 185 190 Val Leu Ile Ala Phe Ala Gln Tyr Leu Gln Gln Cys Pro Phe Glu Asp 195 200 205 His Val Lys Leu Val Asn Glu Val Thr Glu Phe Ala Lys Thr Cys Val 210 215 220 Ala Asp Glu Ser Ala Glu Asn Cys Asp Lys Ser Leu His Thr Leu Phe 225 230 235 240 Gly Asp Lys Leu Cys Thr Val Ala Thr Leu Arg Glu Thr Tyr Gly Glu 245 250 255 Met Ala Asp Cys Cys Ala Lys Gln Glu Pro Glu Arg Asn Glu Cys Phe 260 265 270 Leu Gln His Lys Asp Asp Asn Pro Asn Leu Pro Arg Leu Val Arg Pro 275 280 285 Glu Val Asp Val Met Cys Thr Ala Phe His Asp Asn Glu Glu Thr Phe 290 295 300 Leu Lys Lys Tyr Leu Tyr Glu Ile Ala Arg Arg His Pro Tyr Phe Tyr 305 310 315 320 Ala Pro Glu Leu Leu Phe Phe Ala Lys Arg Tyr Lys Ala Ala Phe Thr 325 330 335 Glu Cys Cys Gln Ala Ala Asp Lys Ala Ala Cys Leu Leu Pro Lys Leu 340 345 350 Asp Glu Leu Arg Asp Glu Gly Lys Ala Ser Ser Ala Lys Gln Arg Leu 355 360 365 Lys Cys Ala Ser Leu Gln Lys Phe Gly Glu Arg Ala Phe Lys Ala Trp 370 375 380 Ala Val Ala Arg Leu Ser Gln Arg Phe Pro Lys Ala Glu Phe Ala Glu 385 390 395 400 Val Ser Lys Leu Val Thr Asp Leu Thr Lys Val His Thr Glu Cys Cys 405 410 415 His Gly Asp Leu Leu Glu Cys Ala Asp Asp Arg Ala Asp Leu Ala Lys 420 425 430 Tyr Ile Cys Glu Asn Gln Asp Ser Ile Ser Ser Lys Leu Lys Glu Cys 435 440 445 Cys Glu Lys Pro Leu Leu Glu Lys Ser His Cys Ile Ala Glu Val Glu 450 455 460 Asn Asp Glu Met Pro Ala Asp Leu Pro Ser Leu Ala Ala Asp Phe Val 465 470 475 480 Glu Ser Lys Asp Val Cys Lys Asn Tyr Ala Glu Ala Lys Asp Val Phe 485 490 495 Leu Gly Met Phe Leu Tyr Glu Tyr Ala Arg Arg His Pro Asp Tyr Ser 500 505 510 Val Val Leu Leu Leu Arg Leu Ala Lys Thr Tyr Glu Thr Thr Leu Glu 515 520 525 Lys Cys Cys Ala Ala Ala Asp Pro His Glu Cys Tyr Ala Lys Val Phe 530 535 540 Asp Glu Phe Lys Pro Leu Val Glu Glu Pro Gln Asn Leu Ile Lys Gln 545 550 555 560 Asn Cys Glu Leu Phe Glu Gln Leu Gly Glu Tyr Lys Phe Gln Asn Ala 565 570 575 Leu Leu Val Arg Tyr Thr Lys Lys Val Pro Gln Val Ser Thr Pro Thr 580 585 590 Leu Val Glu Val Ser Arg Asn Leu Gly Lys Val Gly Ser Lys Cys Cys 595 600 605 Lys His Pro Glu Ala Lys Arg Met Pro Cys Ala Glu Asp Tyr Leu Ser 610 615 620 Val Val Leu Asn Gln Leu Cys Val Leu His Glu Lys Thr Pro Val Ser 625 630 635 640 Asp Arg Val Thr Lys Cys Cys Thr Glu Ser Leu Val Asn Arg Arg Pro 645 650 655 Cys Phe Ser Ala Leu Glu Val Asp Glu Thr Tyr Val Pro Lys Glu Phe 660 665 670 Asn Ala Glu Thr Phe Thr Phe His Ala Asp Ile Cys Thr Leu Ser Glu 675 680 685 Lys Glu Arg Gln Ile Lys Lys Gln Thr Ala Leu Val Glu Leu Val Lys 690 695 700 His Lys Pro Lys Ala Thr Lys Glu Gln Leu Lys Ala Val Met Asp Asp 705 710 715 720 Phe Ala Ala Phe Val Glu Lys Cys Cys Lys Ala Asp Asp Lys Glu Thr 725 730 735 Cys Phe Ala Glu Glu Gly Lys Lys Leu Val Ala Ala Ser Gln Ala Ala 740 745 750 Leu Gly Leu His His His His His His Glu Pro Glu Ala 755 760 765 <210> 104 <211> 708 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 104 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr 20 25 30 Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn 35 40 45 Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe 50 55 60 Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys 65 70 75 80 Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln 85 90 95 Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn 100 105 110 Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu 115 120 125 Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile 130 135 140 Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly 145 150 155 160 Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val Glu 165 170 175 Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys 180 185 190 Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg 195 200 205 Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser 210 215 220 Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile 225 230 235 240 Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu 245 250 255 Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu 260 265 270 Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly 275 280 285 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 290 295 300 Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Gly Gly Pro Gly 305 310 315 320 Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val Glu 325 330 335 Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys 340 345 350 Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg 355 360 365 Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser 370 375 380 Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser 385 390 395 400 Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg 405 410 415 Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp 420 425 430 Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly 435 440 445 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile 450 455 460 Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg 465 470 475 480 Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly 485 490 495 Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser 500 505 510 Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly 515 520 525 Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp 530 535 540 Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe 545 550 555 560 Gly Gly Gly Thr Lys Val Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly 565 570 575 Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Glu Ser Gly 580 585 590 Gly Gly Leu Ala Gln Ala Gly Gly Ser Leu Ser Leu Ser Cys Ala Ala 595 600 605 Ser Gly Phe Thr Val Ser Asn Ser Val Met Ala Trp Tyr Arg Gln Thr 610 615 620 Pro Gly Lys Gln Arg Glu Phe Val Ala Ile Ile Asn Ser Val Gly Ser 625 630 635 640 Thr Asn Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp 645 650 655 Asn Ala Lys Asn Thr Val Tyr Leu Gln Met Asn Asn Leu Lys Pro Glu 660 665 670 Asp Thr Ala Val Tyr Val Cys Asn Arg Asn Phe Asp Arg Ile Tyr Trp 675 680 685 Gly Gln Gly Thr Gln Val Thr Val Ser Ser His His His His His His 690 695 700 Glu Pro Glu Ala 705 <210> 105 <211> 708 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 105 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Glu Ser Gly Gly Gly 20 25 30 Leu Ala Gln Ala Gly Gly Ser Leu Ser Leu Ser Cys Ala Ala Ser Gly 35 40 45 Phe Thr Val Ser Asn Ser Val Met Ala Trp Tyr Arg Gln Thr Pro Gly 50 55 60 Lys Gln Arg Glu Phe Val Ala Ile Ile Asn Ser Val Gly Ser Thr Asn 65 70 75 80 Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala 85 90 95 Lys Asn Thr Val Tyr Leu Gln Met Asn Asn Leu Lys Pro Glu Asp Thr 100 105 110 Ala Val Tyr Val Cys Asn Arg Asn Phe Asp Arg Ile Tyr Trp Gly Gln 115 120 125 Gly Thr Gln Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly 130 135 140 Gly Ser Gly Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys 145 150 155 160 Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu 165 170 175 Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr 180 185 190 Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln 195 200 205 Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala 210 215 220 Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile 225 230 235 240 Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys 245 250 255 Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp 260 265 270 Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro 275 280 285 Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val 290 295 300 Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser 305 310 315 320 Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val 325 330 335 Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly 340 345 350 Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr 355 360 365 Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser 370 375 380 Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser 385 390 395 400 Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly 405 410 415 Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 420 425 430 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Gly Gly Pro 435 440 445 Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val 450 455 460 Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser 465 470 475 480 Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val 485 490 495 Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser 500 505 510 Ser Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile 515 520 525 Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu 530 535 540 Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp 545 550 555 560 Asp Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 565 570 575 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp 580 585 590 Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 595 600 605 Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val 610 615 620 Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr 625 630 635 640 Ser Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 645 650 655 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 660 665 670 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr 675 680 685 Phe Gly Gly Gly Thr Lys Val Glu Ile Lys His His His His His His 690 695 700 Glu Pro Glu Ala 705 <210> 106 <211> 708 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 106 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Glu Ser Gly Gly Gly 20 25 30 Leu Ala Gln Ala Gly Gly Ser Leu Ser Leu Ser Cys Ala Ala Ser Gly 35 40 45 Phe Thr Val Ser Asn Ser Val Met Ala Trp Tyr Arg Gln Thr Pro Gly 50 55 60 Lys Gln Arg Glu Phe Val Ala Ile Ile Asn Ser Val Gly Ser Thr Asn 65 70 75 80 Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala 85 90 95 Lys Asn Thr Val Tyr Leu Gln Met Asn Asn Leu Lys Pro Glu Asp Thr 100 105 110 Ala Val Tyr Val Cys Asn Arg Asn Phe Asp Arg Ile Tyr Trp Gly Gln 115 120 125 Gly Thr Gln Val Thr Val Ser Ser Ser Gly Gly Pro Gly Pro Ala Gly 130 135 140 Met Lys Gly Leu Pro Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys 145 150 155 160 Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu 165 170 175 Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr 180 185 190 Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln 195 200 205 Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala 210 215 220 Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile 225 230 235 240 Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys 245 250 255 Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp 260 265 270 Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro 275 280 285 Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val 290 295 300 Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser 305 310 315 320 Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val 325 330 335 Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly 340 345 350 Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr 355 360 365 Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser 370 375 380 Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser 385 390 395 400 Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly 405 410 415 Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 420 425 430 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Gly Gly Pro 435 440 445 Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val 450 455 460 Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser 465 470 475 480 Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val 485 490 495 Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser 500 505 510 Ser Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile 515 520 525 Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu 530 535 540 Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp 545 550 555 560 Asp Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 565 570 575 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp 580 585 590 Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 595 600 605 Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val 610 615 620 Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr 625 630 635 640 Ser Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 645 650 655 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 660 665 670 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr 675 680 685 Phe Gly Gly Gly Thr Lys Val Glu Ile Lys His His His His His His 690 695 700 Glu Pro Glu Ala 705 <210> 107 <211> 161 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 107 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr 20 25 30 Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn 35 40 45 Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe 50 55 60 Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys 65 70 75 80 Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln 85 90 95 Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn 100 105 110 Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu 115 120 125 Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile 130 135 140 Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr His His His His His 145 150 155 160 His <210> 108 <211> 708 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 108 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr 20 25 30 Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn 35 40 45 Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe 50 55 60 Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys 65 70 75 80 Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln 85 90 95 Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn 100 105 110 Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu 115 120 125 Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile 130 135 140 Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly 145 150 155 160 Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val Glu 165 170 175 Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys 180 185 190 Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg 195 200 205 Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser 210 215 220 Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile 225 230 235 240 Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu 245 250 255 Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu 260 265 270 Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly 275 280 285 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 290 295 300 Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Gly Gly Pro Gly 305 310 315 320 Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val Glu 325 330 335 Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys 340 345 350 Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg 355 360 365 Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser 370 375 380 Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser 385 390 395 400 Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg 405 410 415 Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp 420 425 430 Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly 435 440 445 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile 450 455 460 Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg 465 470 475 480 Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly 485 490 495 Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser 500 505 510 Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly 515 520 525 Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp 530 535 540 Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe 545 550 555 560 Gly Gly Gly Thr Lys Val Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly 565 570 575 Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Glu Ser Gly 580 585 590 Gly Gly Leu Val Gln Ala Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala 595 600 605 Ser Gly Arg Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala 610 615 620 Pro Gly Lys Gln Arg Glu Leu Val Ala Arg Ile Thr Arg Gly Gly Thr 625 630 635 640 Ile Ser Tyr Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp 645 650 655 Asn Ala Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu 660 665 670 Asp Thr Gly Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp 675 680 685 Gly Lys Gly Thr Gln Val Thr Val Ser Ser His His His His His His 690 695 700 Glu Pro Glu Ala 705 <210> 109 <211> 708 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 109 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Glu Ser Gly Gly Gly 20 25 30 Leu Val Gln Ala Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly 35 40 45 Arg Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly 50 55 60 Lys Gln Arg Glu Leu Val Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser 65 70 75 80 Tyr Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala 85 90 95 Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr 100 105 110 Gly Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys 115 120 125 Gly Thr Gln Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly 130 135 140 Gly Ser Gly Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys 145 150 155 160 Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu 165 170 175 Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr 180 185 190 Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln 195 200 205 Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala 210 215 220 Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile 225 230 235 240 Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys 245 250 255 Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp 260 265 270 Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro 275 280 285 Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val 290 295 300 Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser 305 310 315 320 Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val 325 330 335 Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly 340 345 350 Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr 355 360 365 Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser 370 375 380 Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser 385 390 395 400 Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly 405 410 415 Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 420 425 430 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Gly Gly Pro 435 440 445 Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val 450 455 460 Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser 465 470 475 480 Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val 485 490 495 Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser 500 505 510 Ser Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile 515 520 525 Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu 530 535 540 Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp 545 550 555 560 Asp Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 565 570 575 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp 580 585 590 Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 595 600 605 Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val 610 615 620 Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr 625 630 635 640 Ser Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 645 650 655 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 660 665 670 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr 675 680 685 Phe Gly Gly Gly Thr Lys Val Glu Ile Lys His His His His His His 690 695 700 Glu Pro Glu Ala 705 <210> 110 <211> 708 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 110 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Glu Ser Gly Gly Gly 20 25 30 Leu Val Gln Ala Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly 35 40 45 Arg Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly 50 55 60 Lys Gln Arg Glu Leu Val Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser 65 70 75 80 Tyr Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala 85 90 95 Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr 100 105 110 Gly Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys 115 120 125 Gly Thr Gln Val Thr Val Ser Ser Ser Gly Gly Pro Gly Pro Ala Gly 130 135 140 Met Lys Gly Leu Pro Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys 145 150 155 160 Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu 165 170 175 Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr 180 185 190 Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln 195 200 205 Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala 210 215 220 Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile 225 230 235 240 Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys 245 250 255 Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp 260 265 270 Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro 275 280 285 Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val 290 295 300 Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser 305 310 315 320 Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val 325 330 335 Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly 340 345 350 Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr 355 360 365 Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser 370 375 380 Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser 385 390 395 400 Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly 405 410 415 Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 420 425 430 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Gly Gly Pro 435 440 445 Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val 450 455 460 Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser 465 470 475 480 Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val 485 490 495 Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser 500 505 510 Ser Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile 515 520 525 Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu 530 535 540 Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp 545 550 555 560 Asp Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 565 570 575 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp 580 585 590 Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 595 600 605 Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val 610 615 620 Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr 625 630 635 640 Ser Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 645 650 655 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 660 665 670 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr 675 680 685 Phe Gly Gly Gly Thr Lys Val Glu Ile Lys His His His His His His 690 695 700 Glu Pro Glu Ala 705 <210> 111 <211> 579 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 111 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr 20 25 30 Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn 35 40 45 Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe 50 55 60 Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys 65 70 75 80 Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln 85 90 95 Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn 100 105 110 Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu 115 120 125 Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile 130 135 140 Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly 145 150 155 160 Pro Ala Gly Leu Tyr Ala Gln Pro Gly Ser Glu Val Gln Leu Val Glu 165 170 175 Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys 180 185 190 Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg 195 200 205 Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser 210 215 220 Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile 225 230 235 240 Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu 245 250 255 Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu 260 265 270 Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly 275 280 285 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 290 295 300 Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Gly Gly Pro Gly 305 310 315 320 Pro Ala Gly Leu Tyr Ala Gln Pro Gly Ser Glu Val Gln Leu Val Glu 325 330 335 Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys 340 345 350 Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg 355 360 365 Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser 370 375 380 Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser 385 390 395 400 Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg 405 410 415 Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp 420 425 430 Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly 435 440 445 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile 450 455 460 Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg 465 470 475 480 Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly 485 490 495 Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser 500 505 510 Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly 515 520 525 Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp 530 535 540 Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe 545 550 555 560 Gly Gly Gly Thr Lys Val Glu Ile Lys His His His His His His Glu 565 570 575 Pro Glu Ala <210> 112 <211> 579 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 112 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr 20 25 30 Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn 35 40 45 Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe 50 55 60 Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys 65 70 75 80 Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln 85 90 95 Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn 100 105 110 Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu 115 120 125 Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile 130 135 140 Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Pro 145 150 155 160 Gly Gly Pro Ala Gly Ile Gly Pro Gly Ser Glu Val Gln Leu Val Glu 165 170 175 Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys 180 185 190 Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg 195 200 205 Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser 210 215 220 Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile 225 230 235 240 Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu 245 250 255 Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu 260 265 270 Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly 275 280 285 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 290 295 300 Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Gly Gly Pro Pro 305 310 315 320 Gly Gly Pro Ala Gly Ile Gly Pro Gly Ser Glu Val Gln Leu Val Glu 325 330 335 Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys 340 345 350 Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg 355 360 365 Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser 370 375 380 Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser 385 390 395 400 Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg 405 410 415 Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp 420 425 430 Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly 435 440 445 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile 450 455 460 Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg 465 470 475 480 Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly 485 490 495 Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser 500 505 510 Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly 515 520 525 Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp 530 535 540 Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe 545 550 555 560 Gly Gly Gly Thr Lys Val Glu Ile Lys His His His His His His Glu 565 570 575 Pro Glu Ala <210> 113 <211> 579 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 113 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr 20 25 30 Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn 35 40 45 Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe 50 55 60 Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys 65 70 75 80 Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln 85 90 95 Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn 100 105 110 Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu 115 120 125 Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile 130 135 140 Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Ala 145 150 155 160 Leu Phe Lys Ser Ser Phe Pro Pro Gly Ser Glu Val Gln Leu Val Glu 165 170 175 Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys 180 185 190 Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg 195 200 205 Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser 210 215 220 Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile 225 230 235 240 Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu 245 250 255 Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu 260 265 270 Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly 275 280 285 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 290 295 300 Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Gly Gly Pro Ala 305 310 315 320 Leu Phe Lys Ser Ser Phe Pro Pro Gly Ser Glu Val Gln Leu Val Glu 325 330 335 Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys 340 345 350 Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg 355 360 365 Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser 370 375 380 Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser 385 390 395 400 Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg 405 410 415 Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp 420 425 430 Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly 435 440 445 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile 450 455 460 Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg 465 470 475 480 Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly 485 490 495 Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser 500 505 510 Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly 515 520 525 Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp 530 535 540 Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe 545 550 555 560 Gly Gly Gly Thr Lys Val Glu Ile Lys His His His His His His Glu 565 570 575 Pro Glu Ala <210> 114 <211> 581 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 114 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr 20 25 30 Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn 35 40 45 Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe 50 55 60 Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys 65 70 75 80 Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln 85 90 95 Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn 100 105 110 Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu 115 120 125 Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile 130 135 140 Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Pro 145 150 155 160 Leu Ala Gln Lys Leu Lys Ser Ser Pro Gly Ser Glu Val Gln Leu Val 165 170 175 Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser 180 185 190 Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val 195 200 205 Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly 210 215 220 Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr 225 230 235 240 Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser 245 250 255 Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser 260 265 270 Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly 275 280 285 Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 290 295 300 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Gly Gly Pro 305 310 315 320 Pro Leu Ala Gln Lys Leu Lys Ser Ser Pro Gly Ser Glu Val Gln Leu 325 330 335 Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu 340 345 350 Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp 355 360 365 Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp 370 375 380 Ser Ser Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr 385 390 395 400 Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser 405 410 415 Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn 420 425 430 Trp Asp Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser 435 440 445 Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 450 455 460 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 465 470 475 480 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn 485 490 495 Val Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile 500 505 510 Tyr Ser Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly 515 520 525 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 530 535 540 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr 545 550 555 560 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys His His His His His 565 570 575 His Glu Pro Glu Ala 580 <210> 115 <211> 613 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 115 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr 20 25 30 Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn 35 40 45 Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe 50 55 60 Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys 65 70 75 80 Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln 85 90 95 Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn 100 105 110 Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu 115 120 125 Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile 130 135 140 Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Pro 145 150 155 160 Gly Gly Pro Ala Gly Ile Gly Ala Leu Phe Lys Ser Ser Phe Pro Pro 165 170 175 Leu Ala Gln Lys Leu Lys Ser Ser Pro Gly Ser Glu Val Gln Leu Val 180 185 190 Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser 195 200 205 Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val 210 215 220 Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly 225 230 235 240 Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr 245 250 255 Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser 260 265 270 Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser 275 280 285 Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly 290 295 300 Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 305 310 315 320 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Gly Gly Pro 325 330 335 Pro Gly Gly Pro Ala Gly Ile Gly Ala Leu Phe Lys Ser Ser Phe Pro 340 345 350 Pro Leu Ala Gln Lys Leu Lys Ser Ser Pro Gly Ser Glu Val Gln Leu 355 360 365 Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu 370 375 380 Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp 385 390 395 400 Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp 405 410 415 Ser Ser Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr 420 425 430 Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser 435 440 445 Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn 450 455 460 Trp Asp Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser 465 470 475 480 Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 485 490 495 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 500 505 510 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn 515 520 525 Val Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile 530 535 540 Tyr Ser Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly 545 550 555 560 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 565 570 575 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr 580 585 590 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys His His His His His 595 600 605 His Glu Pro Glu Ala 610 <210> 116 <211> 534 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 116 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe 20 25 30 Gly Met Ser Trp Val Arg Gln Gly Gly Gly Gly Gly Leu Asp Gly Asn 35 40 45 Glu Glu Pro Gly Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly 50 55 60 Arg Asp Thr Leu Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser 65 70 75 80 Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg 85 90 95 Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser 100 105 110 Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly 115 120 125 Lys Pro Leu Gly Leu Gln Ala Arg Val Val Gly Gly Gly Gly Thr Gln 130 135 140 Thr Val Val Thr Gln Glu Pro Ser Leu Thr Val Ser Pro Gly Gly Thr 145 150 155 160 Val Thr Leu Thr Cys Ala Ser Ser Thr Gly Ala Val Thr Ser Gly Asn 165 170 175 Tyr Pro Asn Trp Val Gln Gln Lys Pro Gly Gln Ala Pro Arg Gly Leu 180 185 190 Ile Gly Gly Thr Lys Phe Leu Val Pro Gly Thr Pro Ala Arg Phe Ser 195 200 205 Gly Ser Leu Leu Gly Gly Lys Ala Ala Leu Thr Leu Ser Gly Val Gln 210 215 220 Pro Glu Asp Glu Ala Glu Tyr Tyr Cys Thr Leu Trp Tyr Ser Asn Arg 225 230 235 240 Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gly Gly Gly 245 250 255 Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val 260 265 270 Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Lys Leu Ser 275 280 285 Cys Ala Ala Ser Gly Phe Thr Phe Asn Lys Tyr Ala Ile Asn Trp Val 290 295 300 Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Arg Ile Arg Ser 305 310 315 320 Lys Tyr Asn Asn Tyr Ala Thr Tyr Tyr Ala Asp Gln Val Lys Asp Arg 325 330 335 Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr Ala Tyr Leu Gln Met 340 345 350 Asn Asn Leu Lys Thr Glu Asp Thr Ala Val Tyr Tyr Cys Val Arg His 355 360 365 Ala Asn Phe Gly Asn Ser Tyr Ile Ser Tyr Trp Ala Tyr Trp Gly Gln 370 375 380 Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly 385 390 395 400 Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly 405 410 415 Gly Ser Leu Thr Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser Ser 420 425 430 Tyr Ala Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Phe 435 440 445 Val Val Ala Ile Asn Trp Ala Ser Gly Ser Thr Tyr Tyr Ala Asp Ser 450 455 460 Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu 465 470 475 480 Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr 485 490 495 Cys Ala Ala Gly Tyr Gln Ile Asn Ser Gly Asn Tyr Asn Phe Lys Asp 500 505 510 Tyr Glu Tyr Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 515 520 525 His His His His His His 530 <210> 117 <211> 534 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 117 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe 20 25 30 Gly Met Ser Trp Val Arg Gln Gly Gly Gly Gly Gly Leu Asp Gly Asn 35 40 45 Glu Glu Pro Gly Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly 50 55 60 Arg Asp Thr Leu Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser 65 70 75 80 Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg 85 90 95 Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser 100 105 110 Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly 115 120 125 Ser Gly Gly Gly Gly Ser Gly Gly Val Val Gly Gly Gly Gly Thr Gln 130 135 140 Thr Val Val Thr Gln Glu Pro Ser Leu Thr Val Ser Pro Gly Gly Thr 145 150 155 160 Val Thr Leu Thr Cys Ala Ser Ser Thr Gly Ala Val Thr Ser Gly Asn 165 170 175 Tyr Pro Asn Trp Val Gln Gln Lys Pro Gly Gln Ala Pro Arg Gly Leu 180 185 190 Ile Gly Gly Thr Lys Phe Leu Val Pro Gly Thr Pro Ala Arg Phe Ser 195 200 205 Gly Ser Leu Leu Gly Gly Lys Ala Ala Leu Thr Leu Ser Gly Val Gln 210 215 220 Pro Glu Asp Glu Ala Glu Tyr Tyr Cys Thr Leu Trp Tyr Ser Asn Arg 225 230 235 240 Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gly Gly Gly 245 250 255 Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val 260 265 270 Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Lys Leu Ser 275 280 285 Cys Ala Ala Ser Gly Phe Thr Phe Asn Lys Tyr Ala Ile Asn Trp Val 290 295 300 Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Arg Ile Arg Ser 305 310 315 320 Lys Tyr Asn Asn Tyr Ala Thr Tyr Tyr Ala Asp Gln Val Lys Asp Arg 325 330 335 Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr Ala Tyr Leu Gln Met 340 345 350 Asn Asn Leu Lys Thr Glu Asp Thr Ala Val Tyr Tyr Cys Val Arg His 355 360 365 Ala Asn Phe Gly Asn Ser Tyr Ile Ser Tyr Trp Ala Tyr Trp Gly Gln 370 375 380 Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly 385 390 395 400 Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly 405 410 415 Gly Ser Leu Thr Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser Ser 420 425 430 Tyr Ala Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Phe 435 440 445 Val Val Ala Ile Asn Trp Ala Ser Gly Ser Thr Tyr Tyr Ala Asp Ser 450 455 460 Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu 465 470 475 480 Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr 485 490 495 Cys Ala Ala Gly Tyr Gln Ile Asn Ser Gly Asn Tyr Asn Phe Lys Asp 500 505 510 Tyr Glu Tyr Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 515 520 525 His His His His His His 530 <210> 118 <211> 398 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 118 Val Val Gly Gly Gly Gly Thr Gln Thr Val Val Thr Gln Glu Pro Ser 1 5 10 15 Leu Thr Val Ser Pro Gly Gly Thr Val Thr Leu Thr Cys Ala Ser Ser 20 25 30 Thr Gly Ala Val Thr Ser Gly Asn Tyr Pro Asn Trp Val Gln Gln Lys 35 40 45 Pro Gly Gln Ala Pro Arg Gly Leu Ile Gly Gly Thr Lys Phe Leu Val 50 55 60 Pro Gly Thr Pro Ala Arg Phe Ser Gly Ser Leu Leu Gly Gly Lys Ala 65 70 75 80 Ala Leu Thr Leu Ser Gly Val Gln Pro Glu Asp Glu Ala Glu Tyr Tyr 85 90 95 Cys Thr Leu Trp Tyr Ser Asn Arg Trp Val Phe Gly Gly Gly Thr Lys 100 105 110 Leu Thr Val Leu Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 115 120 125 Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln 130 135 140 Pro Gly Gly Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe 145 150 155 160 Asn Lys Tyr Ala Ile Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu 165 170 175 Glu Trp Val Ala Arg Ile Arg Ser Lys Tyr Asn Asn Tyr Ala Thr Tyr 180 185 190 Tyr Ala Asp Gln Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asp Ser 195 200 205 Lys Asn Thr Ala Tyr Leu Gln Met Asn Asn Leu Lys Thr Glu Asp Thr 210 215 220 Ala Val Tyr Tyr Cys Val Arg His Ala Asn Phe Gly Asn Ser Tyr Ile 225 230 235 240 Ser Tyr Trp Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 245 250 255 Gly Gly Gly Gly Ser Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser 260 265 270 Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Thr Leu Ser Cys Ala 275 280 285 Ala Ser Gly Arg Thr Phe Ser Ser Tyr Ala Met Gly Trp Phe Arg Gln 290 295 300 Ala Pro Gly Lys Glu Arg Glu Phe Val Val Ala Ile Asn Trp Ala Ser 305 310 315 320 Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser 325 330 335 Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg 340 345 350 Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Ala Gly Tyr Gln Ile Asn 355 360 365 Ser Gly Asn Tyr Asn Phe Lys Asp Tyr Glu Tyr Asp Tyr Trp Gly Gln 370 375 380 Gly Thr Leu Val Thr Val Ser Ser His His His His His His 385 390 395 <210> 119 <211> 518 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 119 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe 20 25 30 Gly Met Ser Trp Val Arg Gln Gly Gly Gly Gly Gly Leu Asp Gly Asn 35 40 45 Glu Glu Pro Gly Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly 50 55 60 Arg Asp Thr Leu Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser 65 70 75 80 Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg 85 90 95 Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser 100 105 110 Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly 115 120 125 Lys Pro Leu Gly Leu Gln Ala Arg Val Val Gly Gly Gly Gly Thr Gln 130 135 140 Thr Val Val Thr Gln Glu Pro Ser Leu Thr Val Ser Pro Gly Gly Thr 145 150 155 160 Val Thr Leu Thr Cys Ala Ser Ser Thr Gly Ala Val Thr Ser Gly Asn 165 170 175 Tyr Pro Asn Trp Val Gln Gln Lys Pro Gly Gln Ala Pro Arg Gly Leu 180 185 190 Ile Gly Gly Thr Lys Phe Leu Val Pro Gly Thr Pro Ala Arg Phe Ser 195 200 205 Gly Ser Leu Leu Gly Gly Lys Ala Ala Leu Thr Leu Ser Gly Val Gln 210 215 220 Pro Glu Asp Glu Ala Glu Tyr Tyr Cys Thr Leu Trp Tyr Ser Asn Arg 225 230 235 240 Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gly Gly Gly 245 250 255 Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val 260 265 270 Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Lys Leu Ser 275 280 285 Cys Ala Ala Ser Gly Phe Thr Phe Asn Lys Tyr Ala Ile Asn Trp Val 290 295 300 Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Arg Ile Arg Ser 305 310 315 320 Lys Tyr Asn Asn Tyr Ala Thr Tyr Tyr Ala Asp Gln Val Lys Asp Arg 325 330 335 Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr Ala Tyr Leu Gln Met 340 345 350 Asn Asn Leu Lys Thr Glu Asp Thr Ala Val Tyr Tyr Cys Val Arg His 355 360 365 Ala Asn Phe Gly Asn Ser Tyr Ile Ser Tyr Trp Ala Tyr Trp Gly Gln 370 375 380 Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly 385 390 395 400 Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly 405 410 415 Gly Ser Leu Thr Leu Ser Cys Ala Ala Ser Arg Phe Met Ile Ser Glu 420 425 430 Tyr His Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp 435 440 445 Val Ser Thr Ile Asn Pro Ala Gly Thr Thr Asp Tyr Ala Glu Ser Val 450 455 460 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 465 470 475 480 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 485 490 495 Asp Ser Tyr Gly Tyr Arg Gly Gln Gly Thr Gln Val Thr Val Ser Ser 500 505 510 His His His His His His 515 <210> 120 <211> 518 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 120 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe 20 25 30 Gly Met Ser Trp Val Arg Gln Gly Gly Gly Gly Gly Leu Asp Gly Asn 35 40 45 Glu Glu Pro Gly Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly 50 55 60 Arg Asp Thr Leu Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser 65 70 75 80 Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg 85 90 95 Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser 100 105 110 Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly 115 120 125 Ser Gly Gly Gly Gly Ser Gly Gly Val Val Gly Gly Gly Gly Thr Gln 130 135 140 Thr Val Val Thr Gln Glu Pro Ser Leu Thr Val Ser Pro Gly Gly Thr 145 150 155 160 Val Thr Leu Thr Cys Ala Ser Ser Thr Gly Ala Val Thr Ser Gly Asn 165 170 175 Tyr Pro Asn Trp Val Gln Gln Lys Pro Gly Gln Ala Pro Arg Gly Leu 180 185 190 Ile Gly Gly Thr Lys Phe Leu Val Pro Gly Thr Pro Ala Arg Phe Ser 195 200 205 Gly Ser Leu Leu Gly Gly Lys Ala Ala Leu Thr Leu Ser Gly Val Gln 210 215 220 Pro Glu Asp Glu Ala Glu Tyr Tyr Cys Thr Leu Trp Tyr Ser Asn Arg 225 230 235 240 Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gly Gly Gly 245 250 255 Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val 260 265 270 Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Lys Leu Ser 275 280 285 Cys Ala Ala Ser Gly Phe Thr Phe Asn Lys Tyr Ala Ile Asn Trp Val 290 295 300 Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Arg Ile Arg Ser 305 310 315 320 Lys Tyr Asn Asn Tyr Ala Thr Tyr Tyr Ala Asp Gln Val Lys Asp Arg 325 330 335 Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr Ala Tyr Leu Gln Met 340 345 350 Asn Asn Leu Lys Thr Glu Asp Thr Ala Val Tyr Tyr Cys Val Arg His 355 360 365 Ala Asn Phe Gly Asn Ser Tyr Ile Ser Tyr Trp Ala Tyr Trp Gly Gln 370 375 380 Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly 385 390 395 400 Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly 405 410 415 Gly Ser Leu Thr Leu Ser Cys Ala Ala Ser Arg Phe Met Ile Ser Glu 420 425 430 Tyr His Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp 435 440 445 Val Ser Thr Ile Asn Pro Ala Gly Thr Thr Asp Tyr Ala Glu Ser Val 450 455 460 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 465 470 475 480 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 485 490 495 Asp Ser Tyr Gly Tyr Arg Gly Gln Gly Thr Gln Val Thr Val Ser Ser 500 505 510 His His His His His His 515 <210> 121 <211> 382 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 121 Val Val Gly Gly Gly Gly Thr Gln Thr Val Val Thr Gln Glu Pro Ser 1 5 10 15 Leu Thr Val Ser Pro Gly Gly Thr Val Thr Leu Thr Cys Ala Ser Ser 20 25 30 Thr Gly Ala Val Thr Ser Gly Asn Tyr Pro Asn Trp Val Gln Gln Lys 35 40 45 Pro Gly Gln Ala Pro Arg Gly Leu Ile Gly Gly Thr Lys Phe Leu Val 50 55 60 Pro Gly Thr Pro Ala Arg Phe Ser Gly Ser Leu Leu Gly Gly Lys Ala 65 70 75 80 Ala Leu Thr Leu Ser Gly Val Gln Pro Glu Asp Glu Ala Glu Tyr Tyr 85 90 95 Cys Thr Leu Trp Tyr Ser Asn Arg Trp Val Phe Gly Gly Gly Thr Lys 100 105 110 Leu Thr Val Leu Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 115 120 125 Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln 130 135 140 Pro Gly Gly Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe 145 150 155 160 Asn Lys Tyr Ala Ile Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu 165 170 175 Glu Trp Val Ala Arg Ile Arg Ser Lys Tyr Asn Asn Tyr Ala Thr Tyr 180 185 190 Tyr Ala Asp Gln Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asp Ser 195 200 205 Lys Asn Thr Ala Tyr Leu Gln Met Asn Asn Leu Lys Thr Glu Asp Thr 210 215 220 Ala Val Tyr Tyr Cys Val Arg His Ala Asn Phe Gly Asn Ser Tyr Ile 225 230 235 240 Ser Tyr Trp Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 245 250 255 Gly Gly Gly Gly Ser Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser 260 265 270 Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Thr Leu Ser Cys Ala 275 280 285 Ala Ser Arg Phe Met Ile Ser Glu Tyr His Met His Trp Val Arg Gln 290 295 300 Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Thr Ile Asn Pro Ala Gly 305 310 315 320 Thr Thr Asp Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg 325 330 335 Asp Asn Ala Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Lys Pro 340 345 350 Glu Asp Thr Ala Val Tyr Tyr Cys Asp Ser Tyr Gly Tyr Arg Gly Gln 355 360 365 Gly Thr Gln Val Thr Val Ser Ser His His His His His His 370 375 380 <210> 122 <211> 503 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 122 Gln Val Gln Leu Val Glu Ser Gly Gly Ala Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Pro Val Asn Arg Tyr 20 25 30 Ser Met Arg Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Trp Val 35 40 45 Ala Gly Met Ser Ser Ala Gly Asp Arg Ser Ser Tyr Glu Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ala Arg Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Asn Val Asn Val Gly Phe Glu Tyr Trp Gly Gln Gly Thr Gln Val Thr 100 105 110 Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Ser Glu Val Gln Leu 115 120 125 Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu 130 135 140 Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp 145 150 155 160 Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser 165 170 175 Gly Ser Gly Arg Asp Thr Leu Tyr Ala Asp Ser Val Lys Gly Arg Phe 180 185 190 Thr Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn 195 200 205 Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly 210 215 220 Ser Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly 225 230 235 240 Gly Gly Gly Ser Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly 245 250 255 Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Lys Leu Ser Cys Ala Ala 260 265 270 Ser Gly Phe Thr Phe Asn Lys Tyr Ala Ile Asn Trp Val Arg Gln Ala 275 280 285 Pro Gly Lys Gly Leu Glu Trp Val Ala Arg Ile Arg Ser Lys Tyr Asn 290 295 300 Asn Tyr Ala Thr Tyr Tyr Ala Asp Gln Val Lys Asp Arg Phe Thr Ile 305 310 315 320 Ser Arg Asp Asp Ser Lys Asn Thr Ala Tyr Leu Gln Met Asn Asn Leu 325 330 335 Lys Thr Glu Asp Thr Ala Val Tyr Tyr Cys Val Arg His Ala Asn Phe 340 345 350 Gly Asn Ser Tyr Ile Ser Tyr Trp Ala Tyr Trp Gly Gln Gly Thr Leu 355 360 365 Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 370 375 380 Gly Gly Gly Ser Gln Thr Val Val Thr Gln Glu Pro Ser Leu Thr Val 385 390 395 400 Ser Pro Gly Gly Thr Val Thr Leu Thr Cys Ala Ser Ser Thr Gly Ala 405 410 415 Val Thr Ser Gly Asn Tyr Pro Asn Trp Val Gln Gln Lys Pro Gly Gln 420 425 430 Ala Pro Arg Gly Leu Ile Gly Gly Thr Lys Phe Leu Val Pro Gly Thr 435 440 445 Pro Ala Arg Phe Ser Gly Ser Leu Leu Gly Gly Lys Ala Ala Leu Thr 450 455 460 Leu Ser Gly Val Gln Pro Glu Asp Glu Ala Glu Tyr Tyr Cys Thr Leu 465 470 475 480 Trp Tyr Ser Asn Arg Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val 485 490 495 Leu His His His His His His 500 <210> 123 <211> 509 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 123 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe 20 25 30 Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr Leu Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Thr Ile Gly Gly Ser Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Val 115 120 125 Val Gly Gly Gly Gly Thr Gln Thr Val Val Thr Gln Glu Pro Ser Leu 130 135 140 Thr Val Ser Pro Gly Gly Thr Val Thr Leu Thr Cys Ala Ser Ser Thr 145 150 155 160 Gly Ala Val Thr Ser Gly Asn Tyr Pro Asn Trp Val Gln Gln Lys Pro 165 170 175 Gly Gln Ala Pro Arg Gly Leu Ile Gly Gly Thr Lys Phe Leu Val Pro 180 185 190 Gly Thr Pro Ala Arg Phe Ser Gly Ser Leu Leu Gly Gly Lys Ala Ala 195 200 205 Leu Thr Leu Ser Gly Val Gln Pro Glu Asp Glu Ala Glu Tyr Tyr Cys 210 215 220 Thr Leu Trp Tyr Ser Asn Arg Trp Val Phe Gly Gly Gly Thr Lys Leu 225 230 235 240 Thr Val Leu Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 245 250 255 Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro 260 265 270 Gly Gly Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn 275 280 285 Lys Tyr Ala Ile Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu 290 295 300 Trp Val Ala Arg Ile Arg Ser Lys Tyr Asn Asn Tyr Ala Thr Tyr Tyr 305 310 315 320 Ala Asp Gln Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys 325 330 335 Asn Thr Ala Tyr Leu Gln Met Asn Asn Leu Lys Thr Glu Asp Thr Ala 340 345 350 Val Tyr Tyr Cys Val Arg His Ala Asn Phe Gly Asn Ser Tyr Ile Ser 355 360 365 Tyr Trp Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly 370 375 380 Gly Gly Gly Ser Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly 385 390 395 400 Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Thr Leu Ser Cys Ala Ala 405 410 415 Ser Arg Phe Met Ile Ser Glu Tyr His Met His Trp Val Arg Gln Ala 420 425 430 Pro Gly Lys Gly Leu Glu Trp Val Ser Thr Ile Asn Pro Ala Gly Thr 435 440 445 Thr Asp Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp 450 455 460 Asn Ala Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu 465 470 475 480 Asp Thr Ala Val Tyr Tyr Cys Asp Ser Tyr Gly Tyr Arg Gly Gln Gly 485 490 495 Thr Gln Val Thr Val Ser Ser His His His His His His 500 505 <210> 124 <211> 267 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 124 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Glu Val Gln Leu Val Glu Ser Gly Gly Gly 20 25 30 Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly 35 40 45 Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg Gln Ala Pro Gly 50 55 60 Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser Ser Tyr Thr Tyr 65 70 75 80 Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala 85 90 95 Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr 100 105 110 Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp Ala Leu Asp Tyr 115 120 125 Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser 130 135 140 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln 145 150 155 160 Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr 165 170 175 Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly Trp Tyr Gln Gln 180 185 190 Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser Ala Ser Phe Arg 195 200 205 Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp 210 215 220 Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr 225 230 235 240 Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe Gly Gly Gly Thr 245 250 255 Lys Val Glu Ile Lys His His His His His His 260 265 <210> 125 <211> 5 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: Sortase A cleavage site" <220> <221> MOD_RES <222> (3)..(3) <223> Any amino acid <400> 125 Leu Pro Xaa Thr Gly 1 5 <210> 126 <211> 25 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic peptide" <220> <221> SITE <222> (1)..(25) <223> /note="This sequence may encompass 1-5'Gly Gly Gly Gly Ser' repeating units" <400> 126 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 1 5 10 15 Gly Gly Gly Ser Gly Gly Gly Gly Ser 20 25 <210> 127 <211> 20 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic peptide" <220> <221> SITE <222> (1)..(20) <223> /note="This sequence may encompass 1-5'Gly Gly Gly Ser' repeating units" <400> 127 Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser 1 5 10 15 Gly Gly Gly Ser 20 <210> 128 <211> 7 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: Protease-cleavable sequence" <400> 128 Gly Pro Leu Gly Val Arg Gly 1 5 <210> 129 <211> 8 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: Protease-cleavable sequence" <400> 129 Ile Pro Val Ser Leu Arg Ser Gly 1 5 <210> 130 <211> 8 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: Protease-cleavable sequence" <400> 130 Val Pro Leu Ser Leu Tyr Ser Gly 1 5 <210> 131 <211> 10 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: Protease-cleavable sequence" <400> 131 Ser Gly Glu Ser Pro Ala Tyr Tyr Thr Ala 1 5 10 <210> 132 <211> 5 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic peptide" <400> 132 Gly Gly Gly Gly Ser 1 5 <210> 133 <211> 6 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic peptide" <400> 133 Gly Ser Gly Ser Gly Ser 1 5 <210> 134 <211> 13 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic peptide" <400> 134 Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Thr 1 5 10 <210> 135 <211> 15 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic peptide" <400> 135 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 1 5 10 15 <210> 136 <211> 6 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic 6xHis tag" <400> 136 His His His His His His 1 5 <210> 137 <211> 124 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 137 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe 20 25 30 Gly Met Ser Trp Val Arg Gln Gly Gly Gly Gly Gly Leu Asp Gly Asn 35 40 45 Glu Glu Pro Gly Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly 50 55 60 Arg Asp Thr Leu Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser 65 70 75 80 Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg 85 90 95 Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser 100 105 110 Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser 115 120

Claims (39)

The following structural formula:
[A]-[L1]-[B]-[L2]-[D] or [A]-[L1]-[D]-[L2]-[B] or [D]-[L2]-[B ]-[L1]-[A] or [B]-[L2]-[D]-[L1]-[A] or [D]-[L1]-[B]-[L1]-[A] or As a fusion polypeptide of [B]-[L1]-[D]-[L1]-[A],
A is an interleukin 2 (IL-2) polypeptide;
B is a half-life extending element;
L1 and L2 are each independently a polypeptide linker, wherein L1 is a protease-cleavable polypeptide linker and L2 is an optionally protease-cleavable polypeptide linker;
D is an IL-2 blocking moiety; And
The fusion polypeptide has an attenuated IL-2 receptor activating activity, and the IL-2-receptor activating activity of the fusion polypeptide is a poly containing IL-2 polypeptide produced by cleavage of a linker L1 capable of cleavable protease. A fusion polypeptide that is at least about 10 times lower than the IL-2 receptor activating activity of the peptide. a) interleukin 2 (IL-2) polypeptide [A];
b) IL-2 blocking moiety [D]; And
C) a fusion polypeptide comprising at least one of each of the protease-cleavable polypeptide linkers [L],
The IL-2 polypeptide and the IL-2 blocking moiety are operably linked by a protease-cleavable polypeptide linker, and the fusion polypeptide has an attenuated cytokine receptor activation activity, and the cytokine of the fusion polypeptide A fusion polypeptide wherein the kine-receptor activating activity is at least about 10 times lower than the cytokine receptor activating activity of a polypeptide containing a cytokine polypeptide produced by cleavage of a protease cleavable linker. The fusion polypeptide according to claim 1 or 2, wherein the agonist activity of the IL-2 polypeptide containing the fragment of the truncated polypeptide is increased by at least about 50 times compared to the uncleaved fusion polypeptide. The fusion polypeptide according to any one of claims 1 to 3, wherein the agonist activity is assessed using a CTLL-2 proliferation assay, a phospho STAT ELISA or a HEK Blue reporter cell assay. The fusion poly of any one of claims 1 to 4, wherein the uncleaved fusion polypeptide binds to IL-2 receptor alpha (IL-2Rα) in a manner substantially similar to that of naturally occurring IL-2. Peptide. The method of any one of claims 1 to 5, wherein the blocking moiety is IL-2 receptor alpha/beta/gamma (IL-2Rαβγ) and IL- by the IL-2 polypeptide in the uncleaved fusion polypeptide. A fusion polypeptide that inhibits the activation of 2 receptor beta/gamma (IL-2Rβγ). The method of any one of claims 1 to 6, wherein each of the protease-cleavable linker polypeptides is kallikrein, thrombin, chimaase, carboxypeptidase A, cathepsin G, cathepsin L, elastase, PR. -3, granzyme M, calpain, matrix metalloproteinase (MMP), fibroblast activation protein (FAP), ADAM metalloproteinase, plasminogen activator, cathepsin, caspase, tryptase and A fusion polypeptide that independently comprises a sequence that can be cleaved by a protease selected from the group consisting of tumor cell surface proteases. The method of any one of claims 1 to 7, wherein each protease-cleavable polypeptide comprises two or more cleavage sites for the same protease, or two or more cleavage sites cleaved by different proteases, or -At least one of the cleavable polypeptides comprises cleavage sites for at least two different proteases. 9. The fusion polypeptide according to any one of claims 1 to 8, wherein the IL-2 blocking moiety non-covalently binds to the IL-2 polypeptide. The fusion polypeptide according to claim 9, wherein the non-covalent bond is pH dependent. The method of any one of claims 1 to 10, wherein the IL-2 blocking moiety is a ligand binding domain or fragment of a cognate receptor for IL-2, a single domain antibody that binds to an IL-2 polypeptide, Fab or A fusion polypeptide comprising an antibody or antibody fragment (eg, Fab, single domain antibody, scFv) that binds to the scFv or receptor of IL-2. 12. The fusion polypeptide of any one of claims 1-11, wherein the IL-2 blocking moiety is also a half-life extending element. The fusion polypeptide according to any one of claims 1 to 8 or 12, wherein the IL-2 blocking moiety sterically blocks the agonist activity of the IL-2 polypeptide. The fusion polypeptide according to claim 12 or 13, wherein the IL-2 blocking moiety is a human serum albumin or an antigen binding polypeptide that binds to human serum albumin. The fusion polypeptide according to any one of claims 1 to 14, wherein the IL-2 is released by dissociation from the IL-2 blocking moiety after the protease-cleavable polypeptide linker is cleaved by the protease. 16. The fusion polypeptide according to any one of claims 1 to 15, wherein the fusion polypeptide binds to IL-2Rα. 17. The fusion polypeptide of any of the preceding claims, further comprising at least one half-life extending element. The fusion polypeptide according to claim 17, wherein the half-life extending element is human serum albumin, or an antigen binding polypeptide that binds to human serum albumin. The fusion polypeptide of claim 17, wherein the half-life extending element is an immunoglobulin Fc. The fusion polypeptide of claim 13, wherein the IL-2 blocking moiety is human serum albumin, human IgG, humanized IgG, sdAb, Fab and scFv or fragments thereof. The method of any one of claims 1 to 20 o201ne, wherein the IL-2 receptor activation is a standard in vitro Fusion polypeptides, determined using a receptor activation assay and the same amount on a molar basis of IL-2 polypeptide and fusion polypeptide. The fusion of any one of claims 1-21, wherein the protease-cleavable sequence is cleaved by the protease and then the IL-2 is dissociated and released from the IL-2 blocking moiety and/or the half-life extending element. Polypeptide. 20. The fusion polypeptide of any of claims 17-19, wherein the at least one half-life extending element is one half-life extending element or two half-life extending elements. The fusion polypeptide according to any one of claims 1 to 23, wherein L2 is a protease-cleavable polypeptide linker. 25. The fusion polypeptide of any one of claims 1 to 24, wherein L1 is a substrate for a first protease and L2 is a substrate for a second protease. 26. The fusion polypeptide according to any one of claims 2 to 25, further comprising a tumor specific antigen binding peptide. The fusion polypeptide of claim 26, wherein the tumor-specific antigen binding peptide is linked to the IL-2 polypeptide by a non-cleavable linker. The fusion polypeptide of claim 26, wherein the tumor-specific antigen binding peptide is linked to an IL-2 polypeptide, a half-life extending element or an IL-2 blocking moiety by a cleavable linker. 29. The fusion polypeptide of any one of claims 1-28, wherein the serum half-life of the IL-2 polypeptide produced by cleavage of the protease cleavable linker is similar to that of naturally occurring IL-2. The fusion polypeptide according to any one of claims 1 to 29, wherein the IL-2 polypeptide comprises a deletion or substitution of a cystine residue corresponding to Cys125 of SEQ ID NO:. A nucleic acid encoding the polypeptide of any one of claims 1 to 30. A vector comprising the nucleic acid of claim 31. A host cell comprising the vector of claim 32. A method for preparing a pharmaceutical composition comprising culturing the host cell of claim 33 under conditions suitable for expression and collection of the desired polypeptide. A pharmaceutical composition comprising i) an effective amount of the fusion polypeptide of any one of claims 1 to 30 and ii) a pharmaceutically acceptable excipient. A method of treating a tumor, comprising administering an effective amount of the fusion polypeptide of any one of claims 1 to 30 to a subject in need thereof. The fusion polypeptide of any one of claims 1 to 30 for use in medicine. The fusion polypeptide of any one of claims 1 to 30 for use in treating a tumor in a subject in need thereof. A pharmaceutical composition for treating a tumor in a subject in need thereof, comprising the fusion polypeptide of any one of claims 1 to 30 as an active ingredient.

Images